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    ( 254 of 1848 )

United States Patent Application 20180193477
Kind Code A1
Ng; Gordon Yiu Kon ;   et al. July 12, 2018
DRUG-CONJUGATED BI-SPECIFIC ANTIGEN-BINDING CONSTRUCTS

Abstract

Bispecific antigen-binding constructs e.g., antibodies conjugated to drugs (ADCs), which bind CD3 and other cell-surface target antigen such as tumor antigens e.g., CD19, CDH3, HER2, HER3 and EGFR antigens and methods of use are disclosed.

Inventors: Ng; Gordon Yiu Kon; (Vancouver, CA) ; Presta; Leonard G.; (San Francisco, CA) ; Spreter Von Kreudenstein; Thomas; (Vancouver, CA)
Applicant:
Name City State Country Type

Zymeworks Inc.
Vancouver
CA
Family ID: 57756601
Appl. No.: 15/741984
Filed: July 15, 2016
PCT Filed: July 15, 2016
PCT NO: PCT/CA2016/050839
371 Date: January 4, 2018
Related U.S. Patent Documents
Application NumberFiling DatePatent Number
62193056Jul 15, 2015
62193569Jul 16, 2015

Current U.S. Class: 1/1
Current CPC Class: A61K 47/6849 20170801; C07K 2317/56 20130101; C07K 16/46 20130101; A61P 35/00 20180101; C07K 2317/622 20130101; C40B 40/02 20130101; A61K 47/6835 20170801; A61K 47/6869 20170801; C07K 2317/77 20130101; C07K 16/28 20130101; C40B 40/08 20130101; A61K 39/3955 20130101; C07K 16/2863 20130101; C07K 2319/00 20130101; C07K 2317/55 20130101; A61K 47/6817 20170801; A61K 51/1072 20130101; A61K 47/6879 20170801; A61K 2039/505 20130101; C07K 2317/52 20130101; C07K 2317/524 20130101; A61P 35/04 20180101; C07K 2317/24 20130101; C07K 2317/526 20130101; C07K 2317/76 20130101; A61K 39/39558 20130101; C07K 16/32 20130101; C07K 2317/53 20130101; C07K 2317/73 20130101; A61K 51/1027 20130101; A61K 51/1042 20130101; A61K 51/1051 20130101; A61K 2039/572 20130101; A61K 47/6855 20170801; C07K 2317/21 20130101; C07K 2317/92 20130101; C07K 16/2803 20130101; C07K 2317/565 20130101; C07K 2317/94 20130101; A61K 51/103 20130101; C07K 2317/33 20130101; C07K 2317/71 20130101; A61K 45/06 20130101; A61K 47/6811 20170801; C07K 2317/31 20130101; A61P 35/02 20180101; C07K 2317/75 20130101; C07K 16/2809 20130101; A61K 47/6803 20170801
International Class: A61K 47/68 20060101 A61K047/68; A61P 35/00 20060101 A61P035/00; A61P 35/02 20060101 A61P035/02; A61P 35/04 20060101 A61P035/04; C07K 16/28 20060101 C07K016/28; A61K 51/10 20060101 A61K051/10; C07K 16/32 20060101 C07K016/32; A61K 45/06 20060101 A61K045/06; A61K 39/395 20060101 A61K039/395; C07K 16/46 20060101 C07K016/46

Claims


1. A method of killing target cells that express a target antigen on the cell surface comprising contacting the target cells with an effective amount of a drug-conjugated antigen-binding construct in the presence of effector T cells, wherein the drug-conjugated antigen-binding construct comprises a first antigen-binding polypeptide construct that specifically binds a CD3 antigen expressed on the effector T cells, comprising a first heavy chain variable (VH) region and a first light chain variable (VL) region; a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds the target antigen; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and wherein the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

2. A method of killing target cells that express a target antigen on the cell surface in a subject, comprising administering to the subject an effective amount of a drug-conjugated antigen-binding construct comprising a first antigen-binding polypeptide construct comprising a first VH region and a first VL region that specifically binds a CD3 antigen expressed on T cells of the subject; a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds the target antigen; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and wherein the target antigen is not CD3, and wherein the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

3. A method of treating a disease, disorder or condition in a subject in need thereof comprising administering to the subject therapeutically effective amount of a drug-conjugated antigen-binding construct comprising a first antigen-binding polypeptide construct comprising a first VH region and a first VL region that specifically binds a CD3 antigen expressed on T cells of the subject; a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds a disease-associated target antigen; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and wherein the target antigen is not CD3, and wherein the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

4. A composition consisting of a drug-conjugated antigen-binding construct comprising a first antigen-binding polypeptide construct comprising a first VH region, and optionally and a first VL region, that specifically binds a CD3 antigen expressed on a T cell; a second antigen-binding polypeptide construct comprising a second VH region, and optionally a second VL region, that specifically binds a target antigen expressed on a target cell; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and wherein the target antigen is not CD3; and wherein the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

5. The method of any one of claims 1-3 or the composition of claim 4 wherein the first antigen-binding polypeptide construct is a Fab and the second antigen-binding polypeptide is an scFv; or the first antigen-binding polypeptide construct is an scFv and the second antigen-binding polypeptide is a Fab; or the first and second antigen-binding polypeptide constructs are scFvs; or the first and second antigen-binding polypeptide constructs are Fabs.

6. The method or composition of any one of claims 1-5 wherein the antigen-binding construct further comprises a scaffold.

7. The method or composition of claim 6 wherein the scaffold is selected from an Fc, an albumin and a split-albumin.

8. The method or composition of claim 7 wherein the scaffold is an Fc.

9. The method or composition of claim 8, wherein the Fc is a heterodimeric Fc comprising a first Fc polypeptide linked to the first antigen-binding polypeptide construct with or without a first linker and a second Fc polypeptide linked to the second antigen-binding polypeptide construct with or without a second linker.

10. The method or composition of claim 9, wherein the first and second linkers are polypeptides, optionally comprising an IgG1 hinge region.

11. The method or composition of claim 9 or 10, wherein the heterodimeric Fc is a human Fc; and/or is a human IgG1 Fc, IgG.sub.2 or IgG.sub.4 Fc; and/or comprises one or more modifications in at least one of the CH3 domains; and/or comprises one or more modifications in at least one of the CH3 domains that promote formation of the heterodimeric Fc with stability comparable to a wild-type homodimeric Fc; and/or comprises a CH3 region comprising asymmetric amino acid modifications that promote the formation of the heterodimeric Fc and the dimerized CH3 domains having a melting temperature (Tm) of about 68.degree. C. or higher, and/or comprises one or more modifications in at least one of the CH3 domains as described in Table C; and/or further comprises at least one CH2 domain; and/or further comprises at least one CH2 domain comprising one or more modifications; and/or further comprises at least one CH2 domain comprising one or more modifications in at least one of the CH2 domains as described in Table D or Table E; and/or comprises one or more modifications to promote selective binding of Fc-gamma receptors and/or complement; and/or comprises a hinge region with one or more modifications.

12. The method or composition of claim 11, wherein the antigen-binding construct displays reduced Fc gamma receptor binding and reduced associated immune-cell mediated effector activity as compared to a native IgG1 Fc domain.

13. The method or composition of claim 11 or 12 wherein both Fc polypeptides comprise the amino acid modifications L234A, L235A and D265S.

14. The method or composition of any preceding claim wherein the first antigen-binding polypeptide construct is derived from, or competes for binding to CD3 by at least 50%, with, an antibody selected from OKT3; teplizumab; blinatumomab; UCHT1; NI0401; visilizumab; X35-3, VIT3, BMA030 (BW264/56), CLB-T3/3, CRIS7, YTH12.5, F111-409, CLB-T3.4.2, WT31, WT32, SPv-T3b, 11D8, XIII-141, XIII-46, XIII-87, 12F6, T3/RW2-8C.sub.8, T3/RW2-4B6, OKT3D, M-T301, SMC2, SP34 and F101.01; or a humanized version thereof.

15. The method or composition of any preceding claim wherein the first antigen-binding polypeptide construct is species cross-reactive, optionally cross-reactive to non-chimpanzee primates.

16. The method or composition of any preceding claim wherein the first antigen-binding polypeptide construct comprises 6 CDRs comprising the 6 CDRs of wild-type OKT3, the 6 CDRs of the stabilized variant of OKT3, or the 6 CDRs of the humanized variant of OKT3 in Table S1.

17. The method or composition of any preceding claim wherein the first VL region comprises three CDRs with the amino acid sequences of the CDRs of v15192, v 15193, v 15194, v15195 or v17118 specific for CD3, and the first VH region comprises three CDRs with the amino acid sequences of the CDRs of v15192, v 15193, v 15194 or v15195 specific for CD3.

18. The method or composition of any preceding claim wherein the first VL region comprises an amino acid sequence selected from the VL amino acid sequence of the first antigen-binding polypeptide construct of v15192, v 15193, v 15194, v15195, or 17118 specific for CD3 and the first VH region comprises an amino acid sequence selected from the VH amino acid sequence of the first antigen-binding polypeptide construct of v15192, v 15193, v 15194, v15195 or 17118 specific for CD3.

19. The method or composition of any preceding claim wherein the drug is a toxin, a chemotherapeutic agent, a small molecule therapeutic, an immune modulator or a radioisotope.

20. The method or composition of claim 19 wherein the toxin is a microtubule disrupting agent or a DNA modifying agent that is covalently bound to the antigen-binding construct through a linker.

21. The method or composition of claim 20 wherein the microtubule disrupting agent is a maytansinoid, an auristatin, a dolastatin, a tubulysin, or a hemiasterlin.

22. The method or composition of any preceding claim, wherein the drug is a maytansinoid, optionally DM1 or DM4.

23. The method or composition of any preceding claim wherein the drug is an auristatin, optionally MMAE.

24. The method or composition of claim 20 wherein the linker is a non-cleavable linker, optionally SMCC.

25. The method or composition of claim 20 wherein the linker is a cleavable linker, optionally SPBD.

26. The method or composition of any preceding claim wherein the DAR (drug/antigen-binding construct ratio) is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, 17, 18, 19 or 20.

27. The method of any preceding claim wherein the DAR is in the range of 2 to 4.

28. The method or composition of any preceding claim wherein wherein the target antigen is associated with a bacterial disease, a parasitic disease, a viral disease, a cancer, an autoimmune condition or an inflammatory condition.

29. The method or composition of any preceding claim wherein the target antigen is selected from the target antigens in Table LL.

30. The method or composition of any preceding claim wherein the target antigen is selected from CD19, CDH3, HER2, HER3 and EGFR.

31. The method or composition of any preceding claim wherein the target antigen is CD19.

32. The method or composition of claim 31 wherein the second antigen-binding polypeptide construct is specific for CD19 and is derived from, or competes for binding to CD3 by at least 50% with, an antibody selected from the group consisting of 4G7; B4; B43; BU12; CLB-CD19; Leu-12; SJ25-C.sub.1; J4.119, B43, SJ25C.sub.1, FMC63 (IgG.sub.2a) HD237 (IgG.sub.2b), Mor-208, MEDI-551, and MDX-1342.

33. The method or composition of claim 31 wherein the second antigen-binding polypeptide construct comprises 6 CDRs comprising the 6 CDRs of wild-type HD37, the 6 CDRs of the stabilized variant of HD37, or the 6 CDRs of the humanized variant of HD37 in Table S1.

34. The method or composition of claim 31 wherein the second VL region has an amino acid sequences selected from the VL amino acid sequence of the CD19-binding polypeptide construct of v15192, v 15193, v 15194, v15195, or 17118 and the second VH region has an amino acid sequence selected from the VH amino acid sequence of the CD19-binding polypeptide construct of v15192, v 15193, v 15194, v15195 or 17118.

35. The method or composition of claim 31 wherein the second VL region has an amino acid sequence selected from an amino acid sequence that is 80, 85, 90, 95, 96, 97, 98, or 99% identical to the VL amino acid sequence of the CD19-binding polypeptide construct of v15192, v 15193, v 15194, v15195, or 17118 and the second VH region has an amino acid sequence selected from an amino acid sequence that is 80, 85, 90, 95, 96, 97, 98, or 99% identical to the VH amino acid sequence of the CD19-binding polypeptide construct of v15192, v 15193, v 15194, v15195 or 17118.

36. The method or composition of claim 31 wherein the antigen binding construct is selected from (a) variant 6754, 6751, 1853, 10151, 6475, 6749, 10152, 10153, 6476, 5850, 5851, 5852, or 6325, 1661, 1653, 1662, 1660, 1666, 1801, 6747, 10149, 10150, 1380, 12043, 15192, 15193, 15194, 15195 and 17119; or (b) variant having an amino acid sequence at least 80%, 85, 90%, 95%, 96%, 97%, 98%, or 99% identical to variant 6754, 6751, 1853, 10151, 6475, 6749, 10152, 10153, 6476, 5850, 5851, 5852, or 6325, 1661, 1653, 1662, 1660,1666, 1801, 6747, 10149, 10150, 1380, 12043, 15192, 15193, 15194, 15195 or 17119.

37. The method or composition of any one of claims 31-36, wherein the affinity of the second antigen-binding polypeptide construct for CD19 antigen expressed on a B cell is in the range of about 0.5-1, 1-3, 3-5, 5-7, 7-9, 9-11, 11-13, 13-15, 15-17, 17-19 or 19-21 nM, and the affinity of the first antigen-binding polypeptide construct for CD3 expressed on a T cell is in the range of about 5-10, 10-15, 15-20, 20-15, 25-30, 30-35, 35-40, 40-50, 50-55, 55-60, 60-70, 70-80, 80-90 or 90-100 nm, as determined by FACS analysis.

38. The method or composition of claim 31 wherein the antigen-binding construct is selected from variants 15192, 15193, 15194, 15195 and 17119 and is conjugated to DM1 using an SMCC linker.

39. The method or composition of claim 31 wherein the antigen-binding construct is selected from variants 15192, 15193, 15194, 15195 and 17119 and is conjugated to DM4 using an SPBD linker.

40. The method or composition of claim 31 wherein the antigen-binding construct is selected from variants 15192, 15193, 15194, 15195 and 17119 and is conjugated to MMAE using an mc-Val-Cit-PABC linker.

41. The method or composition of any one of claims 1-30 wherein the target antigen is HER2.

42. The method or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct is specific for HER2 and is derived from a HER 2 antibody listed in Table KK.

43. The method or composition of any of one of claims 1-30 wherein the second antigen-binding polypeptide construct comprises the 6 CDRs of the antigen-binding polypeptide construct specific for HER2 of v13792.

44. The method or composition of any one of claims 1-30 wherein the antigen-binding construct is 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical to v13792.

45. The method or composition of any one of claims 1-30 wherein the target antigen is HER3.

46. The method of or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct is specific for HER3 and is derived from a HER3 antibody listed in Table KK.

47. The method or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct comprises the 6 CDRs of the antigen-binding polypeptide construct specific for HER3 of v13790.

48. The method or composition of any one of claims 1-30 wherein the antigen-binding construct is 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical to v13790.

49. The method or composition of any one of claims 1-30 wherein the target antigen is EGFR.

50. The method or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct is specific for EGFR and is derived from an EGFR antibody listed in Table KK.

51. The method or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct comprises the 6 CDRs of the antigen-binding polypeptide construct specific for EGFR of v16371.

52. The method or composition of any one of claims 1-30 wherein the antigen-binding construct is 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical to v16371.

53. The method or composition of any one of claims 1-30 wherein the target antigen is CDH3.

54. The method or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct is specific for CDH3 and is derived from an antibody listed in Table KK.

55. The method or composition of any one of claims 1-30 wherein the second antigen-binding polypeptide construct comprises the 6 CDRs of the second antigen-binding polypeptide construct specific for CDH3 of v13831.

56. The method or composition of any one of claims 1-30 wherein the antigen-binding construct is 80, 85, 90, 95, 96, 97, 98, 99 or 100% identical to v13831.

57. The method of any one of claims 3, and 5-56 wherein the disease, condition or disorder is cancer.

58. The method of any one of claims 3, and 5-40 wherein the cancer is a hematopoietic cancer, leukemia, a lymphoma, a hematological cancer, a B-cell lymphoma, a non-Hodgkin's lymphoma, a cancer non-responsive to at least one of a CD19 lytic antibody, a CD20 lytic antibody and blinatumomab, a cancer cell regressive after treatment with blinatumomab, ALL, CLL, NHL, Mantle Cell Lymphoma, disseminated B cell diseases and metastases of the brain, lung, liver, and/or bone.

59. The method or composition of any one of the preceding claims, wherein the Tm of the CD3-binding polypeptide construct, in an scFv format, is 2, 4, 6, 8, or 10 degrees C. higher than the reference CD3-binding polypeptide constructs teplizumab or OKT3.

60. The method or composition of any one of the preceding claims wherein the killing potency of the drug conjugated antigen-binding construct against target cells bearing the target antigen is at least 1.2, at least 1.5, at least 2, at least 5, at least 10, at least 15, at least 20, at least 30, at least 40, at least 60, at least 70, at least 80, at least 90 or at least 100 times greater than than the killing potency of a reference unconjugated antigen-binding construct.

61. The method or composition of any one of claims 2-60 wherein the killing potency of the drug-conjugated antigen-binding construct against target cells bearing the target antigen is at least 1.5, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50 or more-fold higher than the killing potency on T cells; and/or the administration of the drug-conjugated antigen-binding construct to the subject does not substantially deplete the T cells of the subject.

62. The method or composition of any one of the claims wherein the drug-conjugated antigen-binding construct causes less up-regulation of PD-1 expressing T cells in an in vitro assay than a reference blinatumomab drug-conjugate.

63. The method or composition of any of the claims wherein the affinity of the drug-conjugated antigen-binding construct displays a. at least 2, at least 5, at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90 or at least 100-fold lower affinity for the CD3 antigen than for the target antigen, as measured by SPR; and/or b. an affinity of less than 10 nM for target cells bearing the target antigen and an affinity in the range of 10 nM-500 nM for T cells as measured by FACS; and/or c. higher internalization into target cells than into T cells as measured by an in vitro assay.

64. The method or composition of any one of the preceding claims wherein the antigen-binding construct is capable of killing the target cells by both T-cell redirected killing and by internalization of the drug, as determined by in vitro assays.

65. A pharmaceutical composition comprising the drug-conjugated antigen-binding construct of any one of claims 4-64 and a pharmaceutical carrier.

66. The pharmaceutical composition of claim 65, the carrier comprising a buffer, an antioxidant, a low molecular weight molecule, a drug, a protein, an amino acid, a carbohydrate, a lipid, a chelating agent, a stabilizer, or an excipient.

67. A pharmaceutical composition for use in medicine comprising the drug-conjugated antigen-binding construct of any one of claims 4-64.

68. A pharmaceutical composition for use in treatment of cancer comprising the drug-conjugated antigen-binding construct of any one of claims 4-64.

69. An isolated polynucleotide or set of isolated polynucleotides comprising at least one nucleic acid sequence that encodes the antigen-binding construct according to any one of claims 3-64.

70. The polynucleotide according to claim 69, wherein the polynucleotide or set of polynucleotides is cDNA.

71. A vector or set of vectors comprising one or more of the polynucleotides or sets of polynucleotides according to claim 69 or 70, optionally selected from the group consisting of a plasmid, a viral vector, a non-episomal mammalian vector, an expression vector, and a recombinant expression vector.

72. An isolated cell comprising a polynucleotide or set of polynucleotides according to claim 69 or 70, the cell optionally selected from a hybridoma, a Chinese Hamster Ovary (CHO) cell, or a HEK293 cell.

73. A method of producing the drug-conjugated antigen-binding construct according to any one of claims 3 to 64, comprising culturing a host cell under conditions suitable for expressing the antigen-binding construct wherein the host cell comprises one or more polynucleotides according to any one of claim 69 or 70, purifying the construct, conjugating the construct to a drug, and purifying the drug-conjugated construct.

74. A kit comprising the drug-conjugated antigen-binding construct according to any one of claims 4 to 64 or the pharmaceutical composition of claims 65-68 and instructions for use.

75. An antigen-binding construct that binds to a CD3 epsilon subunit comprising a first antigen binding polypeptide construct comprising a VH region and a VL region wherein the VH region comprises 3 CDRs comprising the amino acid sequences of the VH CDRs of the humanized variant of OKT 3 in Table S1; and the VL region comprises 3 CDRs comprising the amino acid sequences of the VL CDRs of the humanized variant of OKT3 in Table S1.

76. The antigen-binding construct of claim 75 wherein the VH region comprises an amino acid sequence selected from (a) the amino acid sequence of hVH1 or hVH2 in FIG. 4 and (b) an amino acid sequence that is at least 80, 85, 90, 95, 96, 97, 98 or 99% identical to the amino acid sequence of hVH1 or hVH2 in FIG. 4; and the VL region comprises an amino acid sequence selected from (a) the amino acid sequence of hVL1 or hVL2 in FIG. 4 and (b) an amino acid sequence that is at least 80, 85, 90, 95, 96, 97, 98 or 99% identical to the amino acid sequence of hVL1 or hVL2 in FIG. 4.

77. The antigen-binding construct of claim 75 or 76 further comprising a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds a target antigen expressed on a target cell; wherein the first and second antigen-binding polypeptide constructs are operably linked.

78. The antigen-binding construct of claim 77 wherein the target antigen is CD19.

79. The antigen-binding construct of claim 78 wherein the second VH region comprises an amino acid sequence selected from a. the amino acid sequence of hVH2 or hVH3 in FIG. 2 and b. an amino acid sequence that is at least 80, 85, 90, 95, 96, 97, 98 or 99% identical to the amino acid sequence of hVH2 or hVH3 in FIG. 2; and the second VL region comprises an amino acid sequence selected from a. the amino acid sequence of hVL2, hVL2 (D-E) or hVL2 (D-S) in FIG. 2 and b. an amino acid sequence that is at least 80, 85, 90, 95, 96, 97, 98 or 99% identical to the amino acid sequence of hVL2, hVL2 (D-E) or hVL2 (D-S) in FIG. 2.

80. The antigen-binding construct of any of claims 75-79 conjugated to a drug.

81. The method or composition of any one of claims 1-75 wherein the CD3 antigen is a CD3 epsilon subunit of CD3.

82. The method of claim 28 wherein the subject is provided with an additional treatment selected from surgery, radiation, small molecule therapy, hormone therapy, immunotherapy or a combination thereof

83. The method of claim 2 or claim 3 wherein the subject is a human.
Description


SEQUENCE LISTING

[0001] Not applicable.

FIELD OF THE INVENTION

[0002] The field of the invention is drug-conjugated bi-specific antigen-binding constructs, e.g., antibodies, comprising a CD3 antigen-binding polypeptide construct, e.g., a CD3 binding domain and a second antigen-binding polypeptide construct, e.g., a domain that binds a target antigen expressed on a target cell, e.g. a tumor cell.

BACKGROUND OF THE INVENTION

[0003] In the realm of therapeutic proteins, antibodies with their multivalent target binding features are excellent scaffolds for the design of drug candidates. Advancing these features further, designed bi-specific antibodies and other fused multispecific therapeutics exhibit dual or multiple target specificities and an opportunity to create drugs with novel modes of action. The development of such multivalent and multispecific therapeutic proteins with favorable pharmacokinetics and functional activity has been a challenge.

[0004] Bi-specific antibodies capable of targeting T cells to tumor cells have been identified and tested for their efficacy in the treatment of cancers. Blinatumomab is an example of a bi-specific anti-CD3-CD19 antibody in a format called BiTE.TM. (Bi-specific T-cell Engager) that has been identified for the treatment of B-cell diseases such as relapsed B-cell non-Hodgkin lymphoma and chronic lymphocytic leukemia (Baeuerle et al (2009)12:4941-4944) and is FDA approved. T cell engagers directed against other tumor-associated target antigens have also been made, and several have entered clinical trials: AMG110/MT110 EpCAM for lung cancer, gastric cancer and colorectal cancer; AMG211/MEDI565 CEA for gastrointestinal adenocarcinoma; and AMG 212/BAY2010112 PSMA for prostate cancer (see Suruadevara, C. M. et al, Oncoimmunology. 2015 June; 4(6): e1008339).

[0005] The BiTE.TM. format is a bi-specific single chain antibody construct that links variable domains derived from two different antibodies. Blinatumomab, is highly efficacious in B cell acute lymphocytic leukemia (ALL) with an overall response rate of over 80%, but despite the high efficacy many patients relapse shortly after or during the treatment. In addition, T cell engagers have been shown to be less effective in malignancies like chronic lymphocytic leukemia (CLL). There is a need for more efficacious and durable T cell engager therapies.

[0006] T cell engager antigen-binding constructs are described in the following: International application no. PCT/US2013/050411 filed on Jul. 13, 2013 and titled "Bispecific Asymmetric Heterodimers Comprising Anti-CD3 Constructs;" International application no. PCT/US2014/046436 filed on Jul. 11, 2014 and titled "Bispecific CD3 and CD19 Antigen Binding Constructs;" PCT/US2015/011664 filed on Jan. 15, 2015 and titled "Bispecific CD3 and CD19 Antigen Binding Constructs."

SUMMARY OF THE INVENTION

[0007] Described herein is a drug-conjugated antigen-binding construct comprising a first antigen-binding polypeptide construct that specifically binds a CD3 antigen expressed on T cells; and a second antigen-binding polypeptide construct that specifically binds a disease-associated target antigen expressed on a target cell. The first and second antigen-binding polypeptides are operably linked; and the antigen-binding construct is conjugated to a drug, optionally to 2 different drugs. In some embodiments, the drug-conjugated antigen-binding construct displays higher killing potency against target cells in vitro than a reference antigen-binding construct that is not conjugated to a drug. and does not substantially deplete T cells when administered to a subject. The drug-conjugated antigen-binding construct comprises one or more drug molecules. The drug-conjugated antigen binding construct may comprise a heterodimeric Fc comprising a first Fc polypeptide linked to the first antigen-binding polypeptide construct with or without a first linker and a second Fc polypeptide linked to the second antigen-binding polypeptide construct with or without a second linker. In some embodiments, the target antigen is CD19. In some embodiments, the target antigen is CDH3. In some embodiments, the target antigen is HER2. In some embodiments, the target antigen is CDH3. In some embodiments, the target antigen is EGFR. In some embodiments the target antigen is selected from Table LL.

[0008] One aspect of the present disclosure is a method of killing target cells that express a target antigen on the cell surface comprising contacting the target cells with an effective amount of a drug-conjugated antigen-binding construct in the presence of effector T cells, wherein the drug-conjugated antigen-binding construct comprises a first antigen-binding polypeptide construct that specifically binds a CD3 antigen expressed on the effector T cells, comprising a first heavy chain variable (VH) region and a first light chain variable (VL) region; a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds the target antigen; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

[0009] Another aspect of the present disclosure is a method of killing target cells that express a target antigen on the cell surface in a subject, comprising administering to the subject an effective amount of a drug-conjugated antigen-binding construct wherein the drug-conjugated antigen-binding construct comprises a first antigen-binding polypeptide construct that specifically binds a CD3 antigen expressed on the T cells of the subject, comprising a first heavy chain variable (VH) region and a first light chain variable (VL) region; a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds the target antigen; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

[0010] Another aspect of the present disclosure is a method of treating a disease, disorder or condition in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a drug-conjugated antigen-binding construct wherein the drug-conjugated antigen-binding construct comprises a first antigen-binding polypeptide construct that specifically binds a CD3 antigen expressed on the T cells of the subject, comprising a first heavy chain variable (VH) region and a first light chain variable (VL) region; a second antigen-binding polypeptide construct comprising a second VH region and a second VL region that specifically binds the target antigen; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and the target antigen is not CD3; and wherein (a) the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis; and/or (b) the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

[0011] Another aspect of the present disclosure is a composition consisting of a drug-conjugated antigen-binding construct comprising a first antigen-binding polypeptide construct comprising a first VH region, and optionally a first VL region, that specifically binds a CD3 antigen expressed on a T cell; a second antigen-binding polypeptide construct comprising a second VH region, and optionally a second VL region, that specifically binds a target antigen expressed on a target cell; and at least one drug conjugated to the antigen-binding construct; wherein the first and second antigen-binding polypeptide constructs are operably linked; and wherein the target antigen is not CD3; and wherein the target antigen is not CD3; and wherein the antigen-binding construct displays higher affinity to the target antigen than to CD3 as measured by SPR or FACS analysis. In some embodiments embodiment, the antigen-binding construct displays higher killing potency against target cells bearing the target antigen than against T cells, as measured in an in vitro assay.

[0012] Another aspect of the present disclosure is an antigen-binding construct that binds to a CD3 epsilon subunit comprising a first antigen binding polypeptide construct comprising a VH region and a VL region wherein the VH region comprises 3 CDRs comprising the amino acid sequences of the VH CDRs of the humanized variant of OKT 3 in Table S1; and the VL region comprises 3 CDRs comprising the amino acid sequences of the VL CDRs of the humanized variant of OKT3 in Table S1. In one embodiment, the construct comprises a VH region comprising an amino acid sequence selected from the amino acid sequence of hVH1 or hVH2 in FIG. 2 and an amino acid sequence that is at least 80, 85, 90, 95, 96, 97, 98 or 99% identical to the amino acid sequence of hVH1 or hVH2 in FIG. 4; and the VL region comprises an amino acid sequence selected from the amino acid sequence of hVL1 or hVL2 in FIG. 4 and an amino acid sequence that is at least 80, 85, 90, 95, 96, 97, 98 or 99% identical to the amino acid sequence of hVL1 or hVL2 in FIG. 4.

BRIEF DESCRIPTION OF THE FIGURES

[0013] FIG. 1A depicts schematic representations of designs of antigen-binding constructs conjugated to a drug, the drug being depicted by a "star". One binding domain of the antigen-binding constructs binds to a CD3 antigen, and the other binding domain binds to a "target antigen" expressed on a the cell surface of a target cell. Although there is only one "star", the construct may contain multiple drug molecules which can be the same, or different. FIG. 1A(i) shows a representation of an exemplary antigen-binding construct in which both of the antigen-binding domains of the antigen-binding construct are scFvs, with the VH and VL regions of each scFv connected with a polypeptide linker. Each scFv is also connected to one polypeptide chain of a heterodimeric Fc with a hinge polypeptide linker. The two polypeptide chains of the antigen-binding construct are covalently linked together via disulphide bonds (depicted as thick solid lines lines). FIG. 1A(ii) depicts a representation of an exemplary antigen-binding construct similar to 1A(i), except the CD3 binding domain is a Fab and the target antigen binding domain are scFvs. FIG. 1A(iii) depicts a similar antigen-binding construct in which the CD3 binding domain is an scFv and the target antigen binding domain is a Fab. FIG. 1A(iv) depicts a similar antigen-binding construct in which the both the CD3 and target antigen binding domains are Fabs.

[0014] FIG. 1B depicts exemplary embodiments of antigen binding construct drug conjugates (ADCs). FIG. 1B(i) shows a 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (MCC)-DM1 conjugate in which the linker-toxin is conjugated via a lysine residue on the antigen binding construct; FIG. 1B(ii) shows an N-succinimidyl-4-(2-pyridyldithio)butanoate (SPDB)-DM4 conjugate in which the linker-toxin is conjugated via a lysine residue on the antigen binding construct; FIG. 1B(iii) shows a maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-Val-Cit-PABC)-MMAE conjugate in which the linker-toxin is conjugated via a cysteine residue on the antigen binding construct. "Ab" represents the antigen binding construct, which may be any one of the designs shown in FIGS. 1A-1D. "n" represents the number of linker-toxin moieties conjugated to the antigen binding construct and is between 1 and 20.

[0015] FIG. 2 depicts humanized CD19 VL and VH sequences based on the mouse HD37 VL and VH sequences. Three humanized VL sequences have been provided: hVL2, hVL2 (D-E), and hVL2 (D-S). hVL2 (D-E) contains a D to E substitution in CDR L1, while hVL2 (D-S) contains a D to S substitution in CDR L1. Two humanized VH sequences have been provided: hVH2, and hVH3. The CDR sequences are identified by boxes. The CDRs identified in this figure are exemplary only. As is known in the art, the identification of CDRs may vary depending on the method used to identify them. Alternate CDR definitions for the anti-CD19 VL and VH sequences are shown in Table S1. Modifications to humanize these sequences with respect to the wild-type mouse HD37 antibody sequence are denoted by underlining.

[0016] FIG. 3 depicts a table showing the number according to Kabat for the anti-CD19 VH and VL sequences, based on the anti-CD19 HD37 antibody.

[0017] FIG. 4 depicts humanized CD3 VL and VH sequences based on the mouse OKT3 and teplizumab (a known humanized OKT3) sequences. Two VH sequences have been provided: hVH1 and hVH2. Two VL sequences have been provided: hVL1 and hVL2. The CDR sequences are identified by boxes. The CDRs identified in this figure are exemplary only. As is known in the art, the identification of CDRs may vary depending on the method used to identify them. Alternate CDR definitions for the anti-CD3 VL and VH sequences are shown in Table S1. Modifications to these sequences with respect to the wild-type teplizumab antibody sequence are denoted by underlining.

[0018] FIG. 5 depicts a table showing the number according to Kabat for the anti-CD3 VH and VL sequences, based on the anti-CD3 OKT3 antibody.

[0019] FIG. 6 depicts the SEC profile of a parental murine anti-CD3-CD19 antigen-binding construct v6751(left) and a humanized anti-CD3-CD19 antigen-binding construct v15192 (right), showing the greatly enhanced purity of v15192.

[0020] FIG. 7 depicts a DSC thermogram of exemplary humanized anti-CD3-CD19 antigen-binding constructs compared to a parental murine anti-CD3-CD19 antigen-binding construct, showing the increase in Tm of the humanized variants. Variants marked as A and B represent different production batches of the same variant.

[0021] FIG. 8 depicts the binding of a humanized anti-CD3-CD19 antigen-binding construct v15195 to (panel A) Raji CD19+ B cells; (panel B) Jurkat CD3+ T cells. Panel (C) depicts the percentage of T:B cell doublets detected when v15195 is incubated with human peripheral blood mononuclear cells (PBMC). T:B cell doublets were detected as being both CD20+ and CD4+ or CD8+.

[0022] FIG. 9 depicts an exemplary UPLC-SEC profile of an anti CD3-CD19 antigen-binding construct v12043 after conjugation to the toxin DM1 using an SMCC linker.

[0023] FIG. 10 depicts the results of an assay in which selected exemplary anti-CD3-CD19 variants that were conjugated to DM1 or DM4 were tested at various concentrations for their ability to inhibit the growth of (A) Ramos B cells which express CD19, (B) Jurkat T cells, which express CD3 and not CD19, and (C) Raji B cells which express CD19 but not CD3.

[0024] FIG. 11 depicts the effects of various concentrations of unconjugated variant anti-CD3-CD19 variants 12043, v12043-DM1 and v12043-DM4 on (A) Raji cells, (CD19+) (B) CD8+ T cells and (C) CD8+/CD69+ T cells in 72-hour cultures of Raji cells incubated with allogenic peripheral blood mononuclear cells that had been depleted of B cells.

[0025] FIG. 12 depicts the results of a second experiment conducted as in FIG. 11.

[0026] FIG. 13 depicts the effects of various concentrations of DM1-conjugated anti-CD3-CD19 variants variants 6754 and 6751 as well as DM1 conjugated control variants (v891, blinatumomab and v4372 bivalent monospecific anti-CD19 antibody) on (A) Ramos (CD19+) target B cells, (B) CD4+ T cells, and (C) PD-1+ T cells in 72-hour cultures of Ramos cells incubated with allogenic peripheral blood mononuclear cells that had been depleted of B cells.

[0027] FIG. 14 depicts the results of a second experiment conducted as in FIG. 13.

[0028] FIG. 15. Depicts the cytotoxic effect at various concentrations of an exemplary anti-CD3-CD19 antigen-binding construct v6751 conjugated to DM1 on Raji, an ALL cell line, and Ramos, an NHL cell line, Jurkat, a T cell line, and K562, a cell line that expresses neither CD19 nor CD3. Controls antibodies were a monspecific bivalent anti-CD19 antibody conjugated to DM1 huB12, and an isotype non-specific IgG conjugated to DM1.

[0029] FIG. 16 depicts the effects of an exemplary anti-CD3-CD19 antigen-binding construct v15195, v15195 conjugated to DM1 and blinatumomab on Raji cells after 72 hours of culture.

[0030] FIG. 17 depicts the effect of v15195 conjugated to DM1 at various concentrations against various ALL and NHL cell lines: RS4-11, Nalm-6, Daudi, SUDHL-4 and SUDHL-6.

[0031] FIG. 18A depicts the effect of v15195, v15195 conjugated to DM1 and blinatumomab in cultures of Raji cells co-cultured with human PBMC on CD8+ T cells, CD8+/CD69+ T cells and CD8+/CD25+ T cells. FIG. 18B depicts the proliferation observed in cultures of Raji cells co-cultured with human PBMC-B cells with v6751, blinatumomab and OKT3 antibodies.

[0032] FIG. 19 depicts the effects of an exemplary anti-CD3-CD19 antigen-binding construct v6751, v6751 conjugated to DM1 and a control bivalent mono-specific antibody anti-CD19 antibody, huBU12, and huBU12 conjugated to DM1 on various T cell subpopulations in cultures of PBMC-B cells: CD4+. CD8+, CD4+/CD25+ and CD8+/CD25+.

[0033] FIG. 20A depicts the effects of v15195 and v15195 conjugated to DM1 on CD8+ and CD8+/CD25+ T cells in co-cultures of Raji cells with PBMC-B cells. FIG. 20B depicts the level of cytokines IFNg, IL6 and IL10 in the cultures at 72 hours.

[0034] FIG. 21 depicts the effects of exemplary anti-CD3-CD19 antigen-binding construct v15193 and v15193 conjugated to MMAE at various concentrations on CD8+ T cells and target Ramos B cells in co-cultures with PBMC.

[0035] FIG. 22 depicts the effects of a single intravenous administration to humanized mice of varying doses ranging from 0.1 to 1.0 mg/kg of exemplary anti-CD3-CD19 antigen-binding construct v12043 (without drug conjugation), v12043-DM1 and v12043-DM4 on B and T cells counts in humanized NSG mice over a 5-day period after administration.

[0036] FIG. 23 depicts the effects of a single intravenous administration to humanized mice of varying doses ranging from 0.3 to 9.0 mg/kg of exemplary anti-CD3-CD19 antigen-binding construct v15195 conjugated to DM1 on CD3+ T cells in spleen and peripheral blood at 8 days after administration.

[0037] FIG. 24 depicts the internalization into cells of pHAb-labelled exemplary antigen-binding constructs anti-CD3-CD19 v15195, anti-CD3-EGFR v16371, and anti-CD3-CDH3 v13831 and control antibodies (v2171 UCHT1 anti-CD3 monospecific bivalent antibody; and anti-RSV antibody, Synagis). Cell lines tested were Jurkat, A431, SKOV3, HCT-116 and JIMT1.

[0038] FIG. 25 depicts the direct cytotoxicity/growth inhibition in the absence of T cells on target cells lines by exemplary antigen-binding constructs anti-CD3-CDH3 v13831, anti-CD3-HER2 v13792, anti-CD3-HER3 v13790 all conjugated to DM1 in comparison to an non-specific IgG control v6249. Cell lines tested were MCF7, SKOV3, JIMT1 and Jurkat.

[0039] FIG. 26 depicts the effects of exemplary antigen-binding constructs anti-CD3-CDH3 v13831, and anti-CD3-HER2 v13792 and their DM1 conjugates at various concentrations on JIMT1 tumor target cells co-cultured with PBMCs.

[0040] FIG. 27 depicts the T cell proliferation and activation of different T cell subpopulations in co-cultures of JIMT1 tumor target cells and PBMC to which various concentrations of DM1-conjugated or unconjugated anti-CD3-CDH3 v13831 or anti-CD3-HER2 v13792 were added. The level of CD4, CD4+CD69, CD4+CD25, CD8, CD8+CD69, CD8+CD25 positive T cells were evaluated for each construct.

[0041] FIG. 28 depicts the effect of DM1-conjugated and unconjugated anti-CD3-CDH3 in co-cultures of JIMT tumor target cells and PBMC and different effector to target cell ratios.

DETAILED DESCRIPTION OF THE INVENTION

[0042] Described herein are drug-conjugated bispecific antigen-binding constructs e.g. antibodies, often termed antibody-drug conjugates or ADCs. Provided herein are drug-conjugated antigen-binding constructs that bind to a CD3 antigen expressed on T cells and to a second target antigen expressed on the surface of a target cell, for example a tumor cell, a cell responsible for autoimmunity or a cell infected with a pathogen. These drug-conjugated antigen-binding constructs comprise a first antigen-binding domain that specifically binds to the CD3 antigen expressed on T cells, and a second antigen-binding domain that specifically binds to another target antigen expressed on a the surface of a target cell, and at least one drug molecule conjugated to the antigen-binding construct. The first and second antigen-binding domains may be operably linked to each other, or they may each be linked to a scaffold, such as an Fc domain, as further described herein.

[0043] Certain exemplary bispecific antigen-binding constructs used herein to produce ADCs have been shown elsewhere to be able to bridge CD3-expressing T cells with CD19-expressing B cells, with the formation of immunological synapses. These antigen-binding constructs were able to mediate T cell-directed B cell depletion as measured by in vitro and ex vivo assays, and as assessed in an in vivo model of disease.

[0044] In some embodiments described herein anti-CD3-target antigen drug-conjugated antigen-binding constructs are shown to exhibit higher killing potency in depleting target tumor cells in vitro than the same antigen-binding construct that does not comprise a drug. Unexpectedly, several exemplary CD3-target antigen drug-conjugated antigen-binding constructs are shown herein to exhibit high killing potency against target antigen-expressing tumor cells in vitro while at the same time exhibiting low potency against T cells. Additionally, in some embodiments, these ADCs are shown not to significantly deplete circulating T cells in vivo in humanized mice when administered at doses up to 3 mg/kg. In view of the lack of impact on T cells, and without being bound by theory, it appears that CD3-target antigen drug-conjugated antigen-binding constructs may exert their effect on target cells through two distinct mechanisms: T cell-mediated killing, and toxin/small molecule-mediated killing resulting from internalization of the CD3-target antigen drug-conjugated antigen-binding constructs. Hence the anti-CD3-target antigen drug-conjugated antigen-binding constructs described herein may have an added benefit in the treatment of diseases such as cancer over conventional T-cell engager therapeutics, none of which, to our knowledge, have incorporated a toxin or other drug. Additionally the drug-conjugated bispecific antigen-binding constructs that comprise antigen binding domains for CD3 and target antigens have potential in treating diseases other than cancer, such as autoimmune or inflammatory diseases and diseases caused by intracellular pathogens, by combining the mechanisms of T cell- and drug-mediated killing.

[0045] Also described are pharmaceutical compositions comprising the drug-conjugated antigen-binding constructs and methods of treating a disease, disorder or condition e.g., cancer, using the drug-conjugated antigen-binding constructs described herein.

[0046] Described herein are drug-conjugated antigen-binding constructs comprising a first antigen-binding polypeptide construct that specifically binds a CD3 antigen expressed on T cells, and a second antigen-binding polypeptide construct which and specifically binds a target antigen, such as a tumor antigen expressed on the surface of tumor cells. The first and second antigen-binding polypeptide constructs are operably linked, and the antigen-binding construct is conjugated to a drug. The drug-conjugated antigen-binding construct displays higher killing potency against target cells bearing the target antigen in vitro than a reference antigen-binding construct that is not conjugated to a drug.

[0047] The antigen-binding polypeptide constructs may have different formats. In some embodiments, the first and second antigen-binding polypeptides each comprise a Fab or an scFv. In some embodiments the first antigen-binding polypeptide construct is a Fab and the second antigen-binding polypeptide is an scFv. In some embodiments the first antigen-binding polypeptide construct is a scFv and the second antigen-binding polypeptide is an scFv. In other embodiments, the first and second antigen-binding polypeptide constructs may both comprise Fabs or may both comprise scFvs. In certain embodiments, the CD3-binding polypeptide construct is an scFv and the target antigen-binding construct is a Fab.

[0048] In some embodiments, the drug-conjugated antigen-binding construct further comprises an heterodimeric Fc, with a first Fc polypeptide linked to the first antigen-binding polypeptide construct with or without a first linker and a second Fc polypeptide linked to the second antigen-binding polypeptide construct with or without a second linker. As described in detail below, in some embodiments, the heterodimeric Fc comprises a modified CH3 domain comprising asymmetric amino acid modifications that promote the formation of the heterodimeric Fc and the dimerized CH3 domains having a melting temperature (Tm) of about 68.degree. C. or higher. In some embodiments, the asymmetric amino acid modifications are selected from Table C below.

[0049] In some embodiments, the second antigen-binding polypeptide construct comprises the antigen-binding polypeptide construct specific for CD3 derived from an antibody selected from OKT3; Teplizumab.TM. (MGA031, Eli Lilly); blinatumomab; UCHT1; NI0401; visilizumab; X35-3, VIT3, BMA030 (BW264/56), CLB-T3/3, CRIS7, YTH12.5, F111-409, CLB-T3.4.2, WT31, WT32, SPv-T3b, 11D8, XIII-141, XIII-46, XIII-87, 12F6, T3/RW2-8C8, T3/RW2-4B6, OKT3D, M-T301, SP34, SMC2 and F101.01; or a humanized version thereof. Other CD3 binding moieties are possible, and may be made by methods described herein. In some embodiments, the antigen-binding polypeptide construct has the 6 CDRs of wild-type OKT3, or the 6 CDRs of the stabilized variant of OKT3, or a humanized variant of OKT3 in Table S1.

[0050] In some embodiments described herein, the target antigen (cognate antigen--for the second antigen-binding polypeptide construct) is a B cell antigen. In some embodiments, the target antigen is CD19. Thus in some some embodiments wherein the tumor antigen is CD19, the second antigen-binding polypeptide construct has the 6 CDRs of HD37 or the humanized variants of HD37 as shown in Table S1. In some embodiments, the second antigen-binding polypeptide construct comprises the antigen-binding polypeptide construct specific for CD19 derived from an antibody selected from the group consisting of 4G7; B4; B43; BU12; CLB-CD19; Leu-12; SJ25-C1; J4.119, B43, SJ25C1, FMC63 (IgG2a) HD237 (IgG2b), Mor-208, MEDI-551, and MDX-1342.

[0051] In other embodiments, the drug-conjugated antigen binding construct may be any of variants 6754, 6751, 1853, 10151, 6475, 6749, 10152, 10153, 6476, 5850, 5851, 5852, 6325, 1661, 1653, 1662, 1660, 1666, 1801, 6747, 10149, 10150, 1380 or 12043, 151912, 15193, 15194, 15195, 17118 or 17119. conjugated to a drug.

[0052] In many embodiments of drug-conjugated antigen-binding construct having Fcs, there are modifications in the CH2 domain to reduce or eliminate Fc gamma receptor binding and thus they have no associated immune-cell mediated effector activity.

[0053] In some embodiments of a drug-conjugated antigen-binding construct, the affinity for the first antigen-binding polypeptide construct for CD3 is at least 2, 5, 10, 15 or 20-fold lower than and the affinity of the second antigen-binding polypeptide construct for the target antigen, as determined by SPR or FACS analysis.

[0054] Also provided is a method of treating a disease, disorder or condition in a subject, the method comprising administering an effective amount of the drug-conjugated antigen-binding construct of to the subject. In some embodiments, the cancer is a hematopoietic cancer, leukemia, a lymphoma, a hematological cancer, a B-cell lymphoma, a non-Hodgkin's lymphoma, a cancer non-responsive to at least one of a CD19 lytic antibody, a CD20 lytic antibody and blinatumomab, a cancer cell regressive after treatment with blinatumomab, ALL, CLL, NHL, Mantle Cell Lymphoma, disseminated B cell diseases and metastases of the brain, lung, liver, and/or bone. In some embodiments, the tumor is a solid tumor.

[0055] Also provided is a method of depleting target cells in a subject comprising administering to the subject an effective amount of a drug-conjugated antigen-binding polypeptide construct comprising a first antigen-binding polypeptide construct that monovalently and specifically binds to a CD3 antigen expressed on T cells of the subject and a second antigen-binding polypeptide construct that specifically binds to an antigen expressed on the target cells, wherein the first and second antigen-binding polypeptide constructs are operably linked, and wherein the antigen-binding construct is conjugated to a drug. In some embodiments, the tumor cells in the subject are depleted, but the T cells are not substantially depleted. In some embodiments, the administration does not result in up-regulation of PD-1+ (inhibitory) T cells in the subject.

Bi-Specific Antigen-Binding Constructs for Drug Conjugation

[0056] Provided herein are drug-conjugates of bi-specific antigen-binding constructs, e.g., antibodies, that bind CD3 and a second antigen expressed on target cells. The antigen-binding construct itself comprises two antigen-binding polypeptide constructs, e.g., antigen binding domains specifically binding either CD3 or the target antigen. In some embodiments, the target antigen is associated with a tumor, for example CD19, HER2, HER3, CDH3, or EGFR. In some embodiments, the antigen-binding construct is derived from known antibodies or antigen-binding constructs. As described in more detail below, the antigen-binding polypeptide constructs may have the format of a Fab or an scFv (single chain Fv) and may include an Fc.

[0057] In some embodiments, first antigen-binding polypeptide construct (anti-CD3) may comprise a second scFv comprising a second VL, a second scFv linker, and a second VH or it may comprise a Fab comprising a second VL and a second VH. The second scFv may be selected from the group consisting of the OKT3 scFv, a modified OKT3 scFv, an OKT3 blocking antibody scFv, and a modified OKT3 blocking antibody scFv, wherein the OKT3 blocking antibody blocks by 50% or greater the binding of OKT3 to the epsilon subunit of the CD3 antigen. The second antigen-binding polypeptide construct may comprise the antigen-binding polypeptide construct specific for CD3 derived from an antibody selected from OKT3; Teplizumab.TM. (MGA031, Eli Lilly); Micromet, blinatumomab; UCHT1; NI0401; visilizumab; X35-3, VIT3, BMA030 (BW264/56), CLB-T3/3, CRIS7, YTH12.5, F111-409, CLB-T3.4.2, WT31, WT32, SPv-T3b, 11D8, XIII-141, XIII-46, XIII-87, 12F6, T3/RW2-8C8, T3/RW2-4B6, OKT3D, M-T301, SMC2, F101.01 or SP34.

[0058] In some embodiments, for example, the second antigen-binding polypeptide construct (anti-CD19) may comprise an scFv comprising a first VL, a first scFv linker, and a first VH or it may comprise a Fab comprising a first VL and a first VH. The first scFv may be selected from the group consisting of an anti-CD19 antibody HD37 scFv, a modified HD37 scFv, an HD37 blocking antibody scFv, and a modified HD37 blocking antibody scFv, wherein the HD37 blocking antibody blocks by 50% or greater the binding of HD37 to the CD19 antigen. Alternatively, antigen-binding polypeptide constructs (anti-CD19) may comprise the corresponding Fabs. The first antigen-binding polypeptide construct may comprise the antigen-binding polypeptide construct specific for CD19 derived from an antibody selected from the group consisting of 4G7; B4; B43; BU12; CLB-CD19; Leu-12; SJ25-C1; J4.119, B43, SJ25C1, FMC63 (IgG2a) HD237 (IgG2b), Mor-208, MEDI-551, or MDX-1342.

[0059] The heterodimeric Fc comprises first and second Fc polypeptides each comprising a modified CH3 sequence capable of forming a dimerized CH3 domain, wherein each modified CH3 sequence comprises asymmetric amino acid modifications that promote formation of a heterodimeric Fc and the dimerized CH3 domains have a melting temperature (Tm) of about 68.degree. C. or higher. The first Fc polypeptide is linked to the first antigen-binding polypeptide construct with a first hinge linker, and the second Fc polypeptide is linked to the second antigen-binding polypeptide construct with a second hinge linker. In some embodiments, and as described below, the CH2 domain of the Fc is modified to reduce or eliminate binding of the drug-conjugated antigen-binding constructs to Fc receptors.

[0060] The term "antigen-binding construct" refers to any agent, e.g., polypeptide or polypeptide complex capable of binding to an antigen. In some aspects an antigen-binding construct is a polypeptide that specifically binds to an antigen of interest. An antigen-binding construct can be a monomer, dimer, multimer, a protein, a peptide, or a protein or peptide complex; an antibody, an antibody fragment, or an antigen-binding fragment thereof; an scFv and the like. An antigen-binding construct can be a polypeptide construct that is monospecific, bi-specific, or multispecific. In some aspects, an antigen-binding construct can include, e.g., one or more antigen-binding components (e.g., Fabs or scFvs) linked to one or more Fc. Further examples of antigen-binding constructs suitable for use in ADCs are described below and provided in the Examples.

[0061] The term "bi-specific" is intended to include any agent, e.g., an antigen-binding construct, which has two antigen-binding moieties (e.g. antigen-binding polypeptide constructs), each with a unique binding specificity. For example, a first antigen-binding moiety binds to an epitope on a first antigen, and a second antigen-binding moiety binds to an epitope on a second antigen, where the first antigen is different from the second antigen.

[0062] For example, in some embodiments a bi-specific agent may bind to, or interact with, (a) a cell surface target molecule and (b) an Fc receptor on the surface of an effector cell. In another embodiment, the agent may bind to, or interact with (a) a first cell surface target molecule and (b) a second cell surface target molecule that is different from the first cells surface target molecule. In another embodiment, the agent may bind to and bridge two cells, i.e. interact with (a) a first cell surface target molecule on a first call and (b) a second cell surface target molecule on a second cell that is different from the first cell's surface target molecule.

[0063] In some embodiments, the bi-specific antigen-binding construct bridges CD3-expressing T cells with CD19-expressing B cells, with the formation of immunological synapses and/or mediation of T cell directed B cell depletion.

[0064] A monospecific antigen-binding construct refers to an antigen-binding construct with a single binding specificity. In other words, both antigen-binding moieties bind to the same epitope on the same antigen. Examples of monospecific antigen-binding constructs include the anti-CD19 antibody HD37 and the anti-CD3 antibody OKT3.

[0065] An antigen-binding construct can be an antibody or antigen-binding portion thereof. As used herein, an "antibody" or "immunoglobulin" refers to a polypeptide substantially encoded by an immunoglobulin gene or immunoglobulin genes, or fragments thereof, which specifically bind and recognize an analyte (e.g., antigen). The recognized immunoglobulin genes include the kappa, lambda, alpha, gamma, delta, epsilon and mu constant region genes, as well as the myriad immunoglobulin variable region genes. Light chains are classified as either kappa or lambda. The "class" of an antibody or immunoglobulin refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgG.sub.1, IgG.sub.2, IgG.sub.3, IgG.sub.4, IgA.sub.1, and IgA.sub.2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called .alpha., .delta., .epsilon., .gamma., and .mu., respectively.

[0066] An exemplary immunoglobulin (antibody) structural unit is composed of two pairs of polypeptide chains, each pair having one "light" (about 25 kD) and one "heavy" chain (about 50-70 kD). The N-terminal domain of each chain defines a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. The terms variable light chain (VL) and variable heavy chain (VH) refer to these light and heavy chain domains respectively.

[0067] The IgG.sub.1 heavy chain comprised of the VH, CH1, CH2 and CH3 domains respectively from the N to C-terminus. The light chain is comprised of the VL and CL domains from N to C terminus. The IgG.sub.1 heavy chain comprises a hinge between the CH1 and CH2 domains.

[0068] The term "hypervariable region" or "HVR", as used herein, refers to each of the regions of an antibody variable domain which are hypervariable in sequence and/or form structurally defined loops ("hypervariable loops"). Generally, native four-chain antibodies comprise six HVRs; three in the VH (H1, H2, H3), and three in the VL (L1, L2, L3). HVRs generally comprise amino acid residues from the hypervariable loops and/or from the complementarity determining regions (CDRs), the latter being of highest sequence variability and/or involved in antigen recognition. With the exception of CDR1 in VH, CDRs generally comprise the amino acid residues that form the hypervariable loops. Hypervariable regions (HVRs) are also referred to as "complementarity determining regions" (CDRs), and these terms are used herein interchangeably in reference to portions of the variable region that form the antigen-binding regions. This particular region has been described by Kabat et al., U.S. Dept. of Health and Human Services, Sequences of Proteins of Immunological Interest (1983) and by Chothia et al., J Mol Biot 196:901-917 (1987), where the definitions include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or variants thereof is intended to be within the scope of the term as defined and used herein. The exact residue numbers which encompass a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in the art can routinely determine which residues comprise a particular CDR given the variable region amino acid sequence of the antibody.

[0069] The CDR regions of an antibody may be used to construct a binding protein, including without limitation, an antibody, a scFv, a diabody, and the like. In a certain embodiment, the antigen-binding constructs described herein will comprise at least one or all the CDR regions from an antibody. CDR sequences may be used on an antibody backbone, or fragment thereof, and likewise may include humanized antibodies, or antibodies containing humanized sequences. Methods of identifying CDR portions of an antibody are well known in the art. See, Shirai, H., Kidera, A., and Nakamura, H., H3-rules: Identification of CDR-H3 structures in antibodies, FEBS Lett., 455(1):188-197, 1999; and Almagro J C, Fransson, J. Front Biosci. 13:1619-33 (2008).

Antige-Binding Polypeptide Construct--Format

[0070] The bi-specific antigen-binding construct comprises two antigen-binding polypeptide constructs, e.g., antigen binding domains. The format of the antigen-binding polypeptide construct determines the functional characteristics of the bi-specific antigen-binding construct. In one embodiment, the bi-specific antigen-binding construct has an scFv-scFv format, i.e. both antigen-binding polypeptide constructs are scFvs. In another embodiment the antigen-binding construct has an scFv-Fab format. In another embodiment, both antigen-binding polypeptide constructs are Fabs.

[0071] The format "Single-chain Fv" or "scFv" includes the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. In some embodiments, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains. For a review of scFv see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0072] Other antigen-binding polypeptide construct formats include a Fab fragment or sdAb.

[0073] The "Fab fragment" (also referred to as fragment antigen-binding) contains the constant domain (CL) of the light chain and the first constant domain (CH1) of the heavy chain along with the variable domains VL and VH on the light and heavy chains respectively. The variable domains comprise the complementarity determining loops (CDR, also referred to as hypervariable region) that are involved in antigen-binding. Fab' fragments differ from Fab fragments by the addition of a few residues at the carboxy terminus of the heavy chain CH1 domain including one or more cysteines from the antibody hinge region.

[0074] The "Single domain antibodies" or "sdAb" format is an individual immunoglobulin domain. Sdabs are fairly stable and easy to express as fusion partner with the Fc chain of an antibody (Harmsen M M, De Haard H J (2007). "Properties, production, and applications of camelid single-domain antibody fragments". Appl. Microbiol Biotechnol. 77(1): 13-22). In some embodiments an antigen-binding construct provided herein comprises an antigen-binding polypeptide construct that lacks a light chain, thus comprising a single domain antibody.

scFv Format

[0075] The antigen-binding constructs described herein are bi-specific, e.g., they comprise two antigen-binding polypeptide constructs each capable of specific binding to a distinct antigen. In some embodiments, either or both antigen-binding polypeptide construct is in an scFv format. (i.e., antigen-binding domains composed of a heavy chain variable domain and a light chain variable domain, connected with a polypeptide linker). In one embodiment said scFv are human. In another embodiment said scFv molecules are humanized. The scFvs are optimized for protein expression and yield by the modifications described below.

[0076] The scFv can be optimized by changing the order of the variable domains VL and VH in the scFv. In some embodiments of an scFv in a antigen-binding construct described herein, the C-terminus of the light chain variable region may be connected to the N-terminus of the heavy chain variable region, or the C-terminus of the heavy chain variable region may be connected to the N-terminus of the light chain variable region.

[0077] The variable regions may be connected via a linker peptide, or scFv linker, that allows the formation of a functional antigen-binding moiety. The scFv can be optimized for protein expression and yield by changing composition and/or length of the scFv linker polypeptide. Typical peptide linkers comprise about 2-20 amino acids, and are described herein or known in the art. Suitable, non-immunogenic linker peptides include, for example, (G.sub.4S), (SG.sub.4).sub.n, (G.sub.4S).sub.n, G.sub.4(SG.sub.4).sub.n, or G.sub.2(SG.sub.2).sub.n, linker peptides, wherein n is generally a number between 1 and 10, typically between 2 and 4.

[0078] In some embodiments, the scFv linker is selected from Table below:

TABLE-US-00001 TABLE A scFv linker polypeptide sequences CD19 GGGGSGGGGSGGGGS CD3 GGGGSGGGGSGGGGS SSTGGGGSGGGGSGGGGSDI VEGGSGGSGGSGGSGGVD Generic linkers: GGGGSGGGGSGGGGS GGGGSGGGGSGGGGSGGGGS GSTSGGGSGGGSGGGGSS GSTSGSGKPGSGEGSTKG

[0079] The scFv molecule may be optimized for protein expression and yield by including stabilizing disulfide bridges between the heavy and light chain variable domains, for example as described in Reiter et al. (Nat Biotechnol 14, 1239-1245 (1996)). Hence, in one embodiment the T cell activating bi-specific antigen-binding molecule of the invention comprises a scFv molecule wherein an amino acid in the heavy chain variable domain and an amino acid in the light chain variable domain have been replaced by cysteine so that a disulfide bridge can be formed between the heavy and light chain variable domain. In a specific embodiment the amino acid at position 44 of the light chain variable domain and the amino acid at position 100 of the heavy chain variable domain have been replaced by cysteine (Kabat numbering).

[0080] As is known in the art, scFvs can also be stabilized by mutation of CDR sequences, as described in [Miller et al., Protein Eng Des Set. 2010 July; 23(7):549-57; Igawa et al., MAbs. 2011 May-June; 3(3):243-5; Perchiacca & Tessier, Annu Rev Chem Biomol Eng. 2012; 3:263-86.]. One or more of the above noted modifications to the format and sequence of the scFv may be applied to scFvs of the antigen-binding constructs.

[0081] Humanized CD19 VH and VL

[0082] In some embodiments, and in order to further stabilize the antigen-binding constructs described herein, the wild-type sequences of the HD37 anti-CD19 antibody can be modified to generate humanized VH and VL polypeptide sequences. Modifications to both the framework regions and CDRs can be made in order to obtain VH and VL polypeptide sequences to be used in the CD19-binding scFvs and Fabs of the antigen-binding constructs. In some embodiments, the modifications are those depicted in FIG. 2. In some embodiments, the Tm of a humanized anti-CD19 binding domain is higher than than the Tm of an HD37 binding domain. In some embodiments, the Tm of a humanized anti-CD19 binding domain is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 10 degrees C. higher than than the Tm of an HD37 binding domain.

Humanized CD3 VH and VL

[0083] In some embodiments, and in order to further atabilize the antigen-binding constructs described herein the wild-type sequences of the OKT3 anti-CDS3 antibody are modified to generate humanized VH and VL polypeptide sequences. Modifications to both the framework regions and CDRs can be made in order to obtain VH and VL polypeptide sequences to be used in the CD3-binding scFvs and Fabs of the antigen-binding constructs. In some embodiments, the modifications are those depicted in FIG. 4. In some embodiments, the Tm of a humanized anti-CD19 scFv binding domain is higher than the Tm of an OKT3 or teplizumab binding domain. In some embodiments, the Tm of a humanized anti-CD19 scFv binding domain is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 10 degrees C. higher than the Tm of an OKT3 or teplizumab binding domain.

Antige-Binding Polypeptide Construct--Antigens

[0084] The antigen-binding constructs described herein specifically bind a CD3 antigen and a second target antigen.

[0085] As used herein, the term "antigenic determinant" is synonymous with "antigen" and "epitope," and refers to a site (e.g. a contiguous stretch of amino acids or a conformational configuration made up of different regions of non-contiguous amino acids) on a polypeptide macromolecule to which an antigen-binding moiety binds, forming an antigen-binding moiety-antigen complex. An epitope typically includes at least 3, and more usually, at least 5 or 8-10 amino acids in a unique spatial conformation. The epitope may comprise amino acid residues directly involved in the binding and other amino acid residues, which are not directly involved in the binding, such as amino acid residues which are effectively blocked by the specifically antigen binding peptide; in other words, the amino acid residue is within the footprint of the specifically antigen binding peptide. Antibodies that recognize the same epitope can be verified in a simple immunoassay showing the ability of one antibody to block the binding of another antibody to a target antigen.

[0086] "Specifically binds", "specific binding" or "selective binding" means that the binding is selective for the antigen and can be discriminated from unwanted or non-specific interactions. The ability of an antigen-binding construct to bind to a specific antigenic determinant can be measured either through an enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to one of skill in the art, e.g. surface plasmon resonance (SPR) technique (analyzed on a BIAcore instrument) (Liljeblad et al, Glyco J 17, 323-329 (2000)), and traditional binding assays (Heeley, Endocr Res 28, 217-229 (2002)). In one embodiment, the extent of binding of an antigen-binding moiety to an unrelated protein is less than about 10% of the binding of the antigen-binding construct to the antigen as measured, e.g., by SPR.

[0087] In certain embodiments, an antigen-binding construct that binds to the antigen, or an antigen-binding molecule comprising that antigen-binding moiety, has a dissociation constant (K.sub.D) of <1 .mu.M, <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g. 10.sup..about.8 M or less, e.g. from 10.sup..about.8 M to 10.sup.''13 M, e.g., from 10.sup.''9 M to 10.sup.''13 M).

[0088] "Affinity" refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., a receptor) and its binding partner (e.g., a ligand). Unless indicated otherwise, as used herein, "binding affinity" refers to intrinsic binding affinity which reflects a 1:1 interaction between members of a binding pair (e.g., an antigen-binding moiety and an antigen, or a receptor and its ligand). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (K.sub.D), which is the ratio of dissociation and association rate constants (k.sub.off and k.sub.on, respectively). Thus, equivalent affinities may comprise different rate constants, as long as the ratio of the rate constants remains the same. Affinity can be measured by well established methods known in the art, including those described herein. A particular method for measuring affinity is Surface Plasmon Resonance (SPR), or whole cell binding assays with cells that express the antigen of interest.

[0089] "Reduced binding", for example reduced binding to an Fc receptor, refers to a decrease in affinity for the respective interaction, as measured for example by SPR. For clarity the term includes also reduction of the affinity to zero (or below the detection limit of the analytic method), i.e. complete abolishment of the interaction. Conversely, "increased binding" refers to an increase in binding affinity for the respective interaction.

[0090] An "activating T cell antigen" as used herein refers to an antigenic determinant expressed on the surface of a T lymphocyte, particularly a cytotoxic T lymphocyte, which is capable of inducing T cell activation upon interaction with an antigen-binding molecule. Specifically, interaction of an antigen-binding molecule with an activating T cell antigen may induce T cell activation by triggering the signaling cascade of the T cell receptor complex. In a particular embodiment the activating T cell antigen is CD3.

[0091] "T cell activation" as used herein refers to one or more cellular response of a T lymphocyte, particularly a cytotoxic T lymphocyte, selected from: proliferation, differentiation, cytokine secretion, cytotoxic effector molecule release, cytotoxic activity, and expression of activation markers. The T cell activating bi-specific antigen-binding molecules of the invention are capable of inducing T cell activation. Suitable assays to measure T cell activation are known in the art described herein.

[0092] A "target cell antigen" or "target antigen" as used herein refers to an antigenic determinant presented on the surface of a target cell, for example a B cell in a tumor such as a cancer cell or a cell of the tumor stroma. A tumor antigen is a target cell antigen expressed on a tumor cell. In some embodiments, a tumor antigen or may be overexpressed on tumor cells. As used herein, the terms "first" and "second" with respect to antigen-binding moieties etc., are used for convenience of distinguishing when there is more than one of each type of moiety. Use of these terms is not intended to confer a specific order or orientation of the T cell activating bi-specific antigen-binding molecule unless explicitly so stated.

[0093] The term "cross-species binding" or "interspecies binding" or "species cross-reactive" as used herein means binding of a binding domain described herein to the same target molecule in humans and other organisms for instance, but not restricted to non-chimpanzee primates. Thus, "cross-species binding" or "interspecies binding" is to be understood as an interspecies reactivity to the same molecule "X" (i.e. the homolog) expressed in different species, but not to a molecule other than "X". Cross-species specificity of a monoclonal antibody recognizing e.g. human CD3 epsilon, to a non-chimpanzee primate CD3 epsilon, e.g. macaque CD3 epsilon, can be determined, for instance, by FACS analysis. The FACS analysis is carried out in a way that the respective monoclonal antibody is tested for binding to human and non-chimpanzee primate cells, e.g. macaque cells, expressing said human and non-chimpanzee primate CD3 epsilon antigens, respectively. An appropriate assay is shown in the following examples. The above-mentioned subject matter applies mutatis mutandis for the CD19. The FACS analysis is carried out in a way that the respective monoclonal antibody is tested for binding to human and non-chimpanzee primate cells, e.g. macaque cells, expressing said human and non-chimpanzee primate CD3 or CD19 antigens.

CD3

[0094] The antigen-binding constructs described herein specifically bind a CD3 antigen.

[0095] "CD3" or "CD3 complex" as described herein is a complex of at least five membrane-bound polypeptides in mature T-lymphocytes that are non-covalently associated with one another and with the T-cell receptor. The CD3 complex includes the gamma, delta, epsilon, and zeta chains (also referred to as subunits). Non-human monoclonal antibodies have been developed against some of these chains, as exemplified by the murine antibodies OKT3, SP34, UCHT1 or 64.1. (See e.g., June, et al., J. Immunol. 136:3945-3952 (1986); Yang, et al., J. Immunol. 137:1097-1100 (1986); and Hayward, et al., Immunol. 64:87-92 (1988)). Clustering of CD3 on T cells, e.g., by immobilized anti-CD3-antibodies, leads to T cell activation similar to the engagement of the T cell receptor but independent from its clone typical specificity. Most anti-CD3-antibodies recognize the CD3.epsilon.-chain.

[0096] In some embodiments, the anti-CD3 scFv or Fab is an scFV or Fab of a known anti-CD3 antibody, or is derived from, e.g., is a modified version of the scFv or Fab of a known anti-CD3 antibody. Antibodies directed against human CD3 which provide for variable regions (VH and VL) to be employed in the bi-specific antigen-binding construct described herein are known in the art and include OKT3 (ORTHOCLONE-OKT3.TM. (muromonab-CD3). Additional anti-CD3 antibodies include "OKT3 blocking antibodies" that block by 50% or greater the binding of OKT3 to the epsilon subunit of the CD3 antigen. Examples include but are not limited to Teplizumab.TM. (MGA031, Eli Lilly); UCHT1 (Pollard et al. 1987 J Histochem Cytochem. 35(11):1329-38); NI0401 (WO2007/033230); and visilizumab (US25834597).

[0097] In one embodiment, the bi-specific antigen-binding construct comprises a CD3 antigen-binding polypeptide construct which monovalently and specifically binds a CD3 antigen, where the CD3 antigen-binding polypeptide construct is derived from OKT3 (ORTHOCLONE-OKT3.TM. (muromonab-CD3). In one embodiment the bi-specific antigen-binding construct comprises a CD3 antigen-binding polypeptide construct which monovalently and specifically binds a CD3 antigen, the VH and VL regions of said CD3 antigen-binding polypeptide derived from the CD3 epsilon-specific antibody OKT3.

[0098] In some embodiments, the binding affinity of the first antigen binding polypeptide construct specific for the epsilon subunit of CD3 is between about 1 nM to about 100 nM, or between about 20 nM to about 100 nM, or, e.g., greater than 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, or greater than 90 nM.

[0099] The epitope on the CD3 epsilon subunit to which the OKT3 antibody binds is identified by analysis of the crystal structure of the OKT3 bound to CD3 epsilon (Kjer-Nielsen L. et al., (2004) Proc. Natl. Acad. Sci. USA 101: 7675-7680). The polypeptide sequence of CD3 epsilon is provided in the Table below.

TABLE-US-00002 TABLE B CD3 Epsilon sequence Human T-cell MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGI surface TQTPYKVSISGTTVILTCPQYPGSEILWQHN glycoprotein DKNIGGDEDDKNIGSDEDHLSLKEFSELEQS CD3 epsilon GYYVCYPRGSKPEDANFYLYLRARVCENCME subunit, MDVMSVATIVIVDICITGGLLLLVYYWSKNR UniProt ID: KAKAKPVTRGAGAGGRQRGQNKERPPPVPNP P07766 (207 DYEPIRKGQRDLYSGLNQRRI amino acids) (SEQ ID NO: 350)

[0100] Analysis of this structure indicates that the CDRs of the OKT3 antibody, with respect to the sequence in Table B, contact human CD3 epsilon at residues 56-57 (SE), 68-70 (GDE), and 101-107 (RGSKPED). The binding hotspots in these residues are underlined. These residues are considered to be the epitope to which OKT3 binds. Accordingly, the antigen-binding constructs described herein may comprise an antigen-binding polypeptide construct that specifically binds to this epitope.

[0101] Provided herein are antigen-binding constructs comprising at least one CD3 binding polypeptide construct that binds to a CD3 complex on at least one CD3 expressing cell, where in the CD3 expressing cell is a T-cell. In certain embodiments, the CD3 expressing cell is a human cell. In some embodiments, the CD3 expressing cell is a non-human, mammalian cell. In some embodiments, the T cell is a cytotoxic T cell. In some embodiments the T cell is a CD4.sup.+ or a CD8.sup.+ T cell.

[0102] In certain embodiments of the antigen-binding constructs provided herein, the construct is capable of activating and redirecting cytotoxic activity of a T cell to a target cell such as a B cell. In a particular embodiment, said redirection is independent of MHC-mediated peptide antigen presentation by the target cell and and/or specificity of the T cell.

Target Antigens

CD19

[0103] B-cell antigen CD 19 (CD 19, also known as B-cell surface antigen B4, Leu-12; Uniprot ID # PI5391) is a human pan-B-cell surface marker that is expressed from early stages of pre-B cell development through terminal differentiation into plasma cells. CD 19 promotes the proliferation and survival of mature B cells. It associates in a complex with CD21 on the cell surface. It also associates with CD81 and Leu-13 and potentiates B cell receptor (BCR) signaling. Together with the BCR, CD 19 modulates intrinsic and antigen receptor-induced signaling thresholds critical for clonal expansion of B cells and humoral immunity. In collaboration with CD21 it links the adaptive and the innate immune system. Upon activation, the cytoplasmic tail of CD 19 becomes phosphorylated which leads to binding by Src-family kinases and recruitment of PI-3 kinase. It is also expressed on the vast majority of non-Hodgkin's lymphoma (NHL) cells as well as some leukemias.

[0104] Because of their critical role in regulating the immune system, disregulation of B cells is associated with a variety of disorders. B-cell disorders, also referred to herein as B-cell related diseases, are divided into excessive or uncontrolled proliferation (lymphomas, leukemias), and defects of B-cell development/immunoglobulin production (immunodeficiencies).

The amino acid sequence of CD19 is as follows:

TABLE-US-00003 MPPPRLLFFLLFLTPMEVRPEEPLVVKVEEGDNAVLQCLKGTSDGP TQQLTWSRESPLKPFLKLSLGLPGLGIHMRPLAIWLFIFNVSQQMG GFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKN RSSEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSL SQDLTMAPGSTLWLSCGVPPDSVSRGPLSWTHVHPKGPKSLLSLEL KDDRPARDMWVMETGLLLPRATAQDAGKYYCHRGNLTMSFHLEITA RPVLWHWLLRTGGWKVSAVTLAYLIFCLCSLVGILHLQRALVLRRK RKRMTDPTRRFFKVTPPPGSGPQNQYGNVLSLPTPTSGLGRAQRWA AGLGGTAPSYGNPSSDVQADGALGSRSPPGVGPEEEEGEGYEEPDS EEDSEFYENDSNLGQDQLSQDGSGYENPEDEPLGPEDEDSFSNAES YENEDEELTQPVARTMDFLSPHGSAWDPSREATSLGSQSYEDMRGI LYAAPQLRSIRGQPGPNHEEDADSYENMDNPDGPDPAWGGGGRMGT WSTR.

[0105] In some embodiments, the antigen-binding constructs described herein include an antigen-binding polypeptide construct that binds to a CD19 antigen (anti-CD19 scFv or Fab).

[0106] In some embodiments, the anti-CD19 scFv or Fab is an scFv or Fab of a known anti-CD19 antibody, or is derived from, e.g., is a modified version of the scFv or Fab of a known anti-CD19 antibody. Antibodies directed against CD19 which provide for variable regions (VH and VL) to be employed in the bi-specific antigen-binding construct described herein are known in the art and include HD37, provided by the HD37 hybridoma (Pezzutto (1997), J. Immunol. 138, 2793-9). Additional anti-CD19 antibodies include "HD37 blocking antibodies" that block by 50% or greater the binding of HD37 to the CD19 antigen. Examples include but are not limited to HD237 (IgG2b) (Fourth International Workshop on Human Leukocyte Differentiation Antigens, Vienna, Austria, 1989; and Pezzutto et al., J. Immunol., 138(9):2793-2799 (1987)); 4G7 (Meecker (1984) Hybridoma 3, 305-20); B4 (Freedman (1987) Blood 70, 418-27); B43 (Bejcek (1995) Cancer Res. 55, 2346-51) and Mor-208 (Hammer (2012) Mabs4:5, 571-577).

[0107] In one embodiment said VH(CD19) and VL(CD19) regions (or parts, like CDRs, thereof) are derived from the anti-CD19 antibody HD37, provided by the HD37 hybridoma (Pezzutto (1997), J. Immunol. 138, 2793-9).

[0108] In some embodiments, the binding affinity of the second antigen-binding polypeptide construct for the target antigen is between about 0.1 nM to about 10 nM or less than 5.0, 4.0, 3.0, 2.0, 1.0, 0.9, 0.09, 0.9, 0.7, 0.6, 0.5, 0.4, 0.3, or less than 0.2 nM. In some embodiments, the binding affinity of the second antigen-binding polypeptide construct to CD19 on the surface of CD19+ target cells is in the range of 0.1 to 0.5, 0.5-1, 1-3, 3-5, 5-7, 7-9, 9-11, 11-13, 13-15, 15-17, 17-19 or 19-21 nM as measured by FACS analysis.

[0109] In certain embodiments, the antigen-binding polypeptide construct is an scFv or Fab construct that binds CD19 on a B cell. In some embodiments the scFv or Fab construct is mammalian. In one embodiment said scFv or Fab construct is human. In another embodiment said scFv or Fab construct is humanized. In yet another embodiment said scFv or Fab construct comprises at least one of human heavy and light chain variable regions.

[0110] In certain embodiments, the antigen-binding polypeptide construct exhibits cross-species binding to a least one antigen expressed on the surface of a B cell. In some embodiments, the antigen-binding polypeptide construct of an antigen-binding construct described herein bind to at least one of mammalian CD19. In certain embodiments, the CD19 antigen-binding polypeptide construct binds a human CD19.

CDH3

[0111] In some embodiments, a drug-conjugated antigen-binding construct may have an antigen-binding polypeptide construct directed against CDH3. CDH3, also known as CADH3; cadherin 3, type 1 or P-cadherin (Uniprot ID # P22223) is a member of the cadherin family of cell adhesion proteins that preferentially interact with themselves in a homophilic manner in cell-cell adhesion. CDH3 overexpression is associated with several types of cancer. In some embodiments, anti-CDH3 antibodies in Table KK are used to derive antigen-binding polypeptide constructs specific for CDH3.

TABLE-US-00004 TABLE KK Antibody Patent/paper reference: anti-HER2 trastuzumab PCT/US1998/026266; Baselga J., et al, 1998, Cancer Res., 58: 2825-31 pertuzumab PCT/US2005/025084; DeGrendele H., 2003, Clin Prostate Cancer, 2: 143-5 ertumaxomab PCT/EP2008/001551; Kiewe P, et al, 2006, Clin. Cancer Res., 12: 3085-91 margetuximab PCT/US2009/038201 XMT-1522 PCT/US2015/036431 MIL5_scFv Qiao C, et al., 2013, J. Biomol. Struct. Dyn., 31: 511-21 7C2, 7F3 U.S. Pat. No. 14,511,604 anti-HER3 seribantumab PCT/US2008/002119; Schoeberl B., et al., 2009, Sci Signal, 2: ra31 patritumab PCT/EP2006/012632; LoRusso P., et al, 2013, Clin Cancer Res., 19: 3078-87 elgemtumab PCT/EP2011/064407; Garrett J T., et al., 2013, Cancer Res., 73: 6013-23 lumretuzumab PCT/EP2010/070062; Mirschberger C., et al., 2013, Cancer Res., 73: 5183-94 KTN3379 PCT/US2012/066038; Lee S., et al., 2015, Proc Natl Acad Sci USA., 112: 13225-30 15D5 and 1D9 PCT/US2011/050322 REGN1400 PCT/US2012/056446 anti-EGFR cetuximab PCT/US1996/009847; Prewett M., et al., 1996, J Immunother Emphasis Tumor Immunol, 19: 419-27 panitumumab PCT/US2003/015734; Yang X D, et al., 2001, Crit Rev Oncol Hematol., 38: 17-23 nimotuzumab PCT/CA2012/050034; Spicer J., 2005, Curr. Opin. Mol. Ther., 7: 182-91 necitumumab PCT/US2005/009583; Lu D., et al., 2005, J. Biol. Chem., 280: 19665-7 zalutumumab PCT/US2002/018748; Lammerts van Bueren J J., et al., 2008, Proc. Natl. Acad. Sci. U.S.A., 105: 6109-14 matuzumab PCT/EP2002/001687; Vanhoefer U., et al., 2004, J. Clin. Oncol., 22: 175-84 imgatuzumab PCT/IB2006/000238; Gerdes C A., et al., 2013, Clin Cancer Res., 19: 1126-38 depatuxizumab PCT/US2007/019988; Gan H K., et al., 2007, J. Biol. Chem., 282: 2840-50 anti-CDH3 FF-21101 PCT/JP2010/057694 Oncotherapy Clone #6 PCT/JP2007/054374 PF-03732010 PCT/IB2006/001053; 2010, Zhang C C., et al., Clin. Cancer Res., 16: 5177-88 PCA062 PCT/IB2015/058801 PF-06671008 PCT/IB2015/054829

HER2, HER3 and EGFR

[0112] HER2, HER3 and EGFR are a HER receptors. A "HER receptor" is a receptor protein tyrosine kinase which belongs to the human epidermal growth factor receptor (HER) family and includes EGFR, HER2, HER3 and HER4 receptors. A HER receptor will generally comprise an extracellular domain, which may bind an HER ligand; a lipophilic transmembrane domain; a conserved intracellular tyrosine kinase domain; and a carboxyl-terminal signaling domain harboring several tyrosine residues which can be phosphorylated. HER2, HER3 and EGFR are overexpressed in numerous types of cancer. In some embodiments, the anti-HER2, anti-HER3 and anti-EGFR antibodies in Table KK are used to derive antigen-binding polypeptide constructs.

Other Target Antigens

[0113] In some embodiments, the drug-conjugated antigen-binding construct comprises a second antigen-binding polypeptide construct that is specific for one of the target antigens provided in Table LL. In some embodiments, the target antigen is a pathogen-derived antigen. In an embodiment, the target antigen is a viral antigen. In some embodiments, the target antigen is a fungal antigen. In some embodiments, the target antigen is bacterial. In some embodiments, the target antigen is a parasite antigen. In some embodiments the target antigen is associated with a hematological cancer. In some embodiments, the target antigen is expressed on a solid tumor. In some embodiments, the target antigen is associated with an autoimmune disease.

TABLE-US-00005 TABLE LL Target Antigens Viral targets Family Genus Virus Retroviridae Lentivirus human immunodeficiency virus Papillomaviridae Many Human papilloma virus Paramyxoviridae Pneumovirus Human respiratory syncytial virus Filoviridae Ebolavirus Ebola virus Coronaviridae Betacoronavirus SARS coronavirus Orthomyxoviridae Influenza A, B, C Influenza Hepadnaviridae Orthohepadnavirus Hepatitis B virus Flaviviridae Hepacivirus Hepatitis C virus Flaviviridae Flavivirus Zika virus Flaviviridae Flavivirus Dengue virus Flaviviridae Flavivirus West Nile Virus Herpesviridae Simplexvirus Herpes simplex virus Herpesviridae Lymphocryptovirus Epstein-Barr Virus Herpesviridae Varicellovirus Varicella-Zoster virus Herpesviridae Cytomegalovirus Cytomegalovirus Bacterial/fungal Targets Family Genus Species Brucellaceae Brucella B. melitensis Chlamydiaceae Chlamydia C. trachomatis Chlamydiaceae Chlamydophila C. pneumoniae Clostridiaceae Clostridium C. difficile Coxiellaceae Coxiella C. burnetii Legionellaceae Legionella L. pneumophila (many more) Listeriaceae Listeria L. monocytogenes Mycobacteriaceae Mycobacterium M. tuberculosis, M. leprae Neisseriaceae Neisseria N. gonorrhoeae, N. meningitidis Rickettsiaceae Rickettsia Numerous species in three groups: Spotted fever (R. rickettsii) Typhus (R. prowazekii) and Scrub-typhus (Orientia tsutsugamushi) Enterobacteriaceae Salmonella S. bongori, S. enterica Enterobacteriaceae Shigella S. boydii, S. dysenteriae, S. flexneri, S. sonnei Enterobacteriaceae Yersinia Y. pestis, Y. pseudotuberculosis Tremellaceae Cryptococcus C. neoformans Trichocomaceae Aspergillus Aspergillus spp Parasitic Targets Family Genus Species Cryptosporidiidae Cryptosporidium C. parvum Plasmodium Plasmodium P. falciparum, P. vivax, P. ovale, and P. malariae Trypanosomatidae Leishmania L. donovani (~20 species infect humans) Sarcocystidae Toxoplasma T. gondii Trypanosomatidae Trypanosoma T. cruzi, T. brucei Human targets Gene ID Uniprot ID Disease Association Cancer Hemooncology CD8 P10966 T cell activation? CD19 P15391 B-cell malignancies, autoimmune disease CD20 P11836 Chronic Lymphocytic Leukemia, Non-Hodgkin's Lymphoma, Rheumatoid Arthritis CD22 P20273 Non-Hodgkin's Lymphoma, B-cell malignancies CD30 P28908 Anaplastic Large Cell Lymphoma Hematologic malignancies Hodgkin Lymphoma CD33 P20138 Acute myeloid leukemia CD37 P11049 Acute myeloid leukemia Chronic Lymphocytic Leukemia Non- Hodgkin's Lymphoma CD38 P28907 Hematologic malignancies, Multiple Myeloma CD44v6 P16070 Squamous cell carcinoma, Hematologic malignancies CD74 P04233 Chronic Lymphocytic Leukemia Multiple Myeloma CD79b P40259 Non-Hodgkin's Lymphoma, Systemic lupus erythematosus CD133 O43490 Acute lymphoblastic leukemia Acute myeloid leukemia CD138 P18827 Multiple Myeloma IL-3R.alpha. P26951 Acute myeloid leukemia, Hodgkin Lymphoma BCMA Q02223 B-cell malignancies, Multiple Myeloma CLEC12A Q5QGZ9 Acute myeloid leukemia FLT3 P36888 Acute myeloid leukemia ROR 1 Q01973 Chronic Lymphocytic Leukemia, B- cell malignancies Solid tumor CD70 (CD27L) P32970 Renal Cell Carcinoma, Autoimmune Diseases, Cancer, Inflammatory Diseases CD117 P10721 Inflammatory Diseases, Cancer, Acute myeloid leukemia CD56 P13591 Multiple Myeloma, Solid Tumors CD98 P08195 head and neck squamous cell carcinoma cells with stem cell properties Notch 1 P46531 solid tumors- broad indications Notch 2 Q04721 solid tumors- broad indications Notch 3 Q9UM47 solid tumors- broad indications Notch 4 Q99466 solid tumors- broad indications DL44 Q9NR61 solid tumors- broad indications PSMA Q04609 Prostate Cancer PSA P07288 Prostate Cancer PSCA O43653 Prostate Cancer STEAP1 Q9UHE8 Prostate Cancer, Multiple Others CEACAM4 O75871 Colorectal Cancer CEACAM5 P06731 Colorectal Cancer, Pancreatic Cancer, Gastric Cancer alpha-V integrin P06756 melanoma, glioma, ovarian, and breast cancer EphA2 P29317 solid tumors Epha10 Q5JZY3 Breast Cancer EpCAM P16422 solid tumors Cadherin-19 J3KTP3 Melanoma P-cadherin P22223 solid tumors Nectin-4 Q96NY8 Metastatic Urothelial Cancer Glypican 3 P51654 Liver Cancer EGFR/EGFRvIII P00533 solid tumors- broad indications VEGFR P17948 endothelial cell - solid tumor HER2/neu P04626 Breast Cancer, Head and Neck Cancer, ovarian, prostate Her3 P21860 Solid tumors- IGF1R P08069 solid tumors and hematological malignancies c-MET P08581 Solid tumors folate receptor alpha P15328 Ovarian Cancer folate receptor beta P14207 Acute myeloid leukemia, Ovarian Cancer Endothelin B receptor P24530 Melanoma TF (Tissue Factor) P13726 Pancreatic Cancer, Acute Lung Injury, Inflammatory Diseases MSLN Q13421 Mesothelioma, Breast Cancer, Ovarian cancer ENPP3 014638 Liver Cancer, Renal Cell Carcinoma TPBG Q13641 Non-Small Cell Lung cancer, Renal Cell Carcinoma FAP Q12884 Stromal Targeting, Colorectal Cancer HMW-MAA Q6UVK1 Melanoma, Breast Cancer A33 Q99795 Colorectal Cancer B7-H3 Q5ZPR3 Solid tumors B7-H4 Q7Z7D3 Solid tumors GPNMB Q14956 Breast Cancer, Melanoma, CFC1B P0CG36 Solid tumors TACSTD (Trop2) P09758 Breast Cancer, Gastric Cancer, Pancreatic Cancer TAG-72 Q9XVS1 Prostate, Breast, Colon, Lung, and Pancreatic cancers TIM-3 Q8TDQ0 Immune Checkpoint, Cancer, Autoimmunity, Inflammation Guanylyl cyclase C P25092 Pancreatic Cancer (GCC)/GUCY2C SLC44A4 Q53GD3 Pancreatic Cancer, Prostate Cancer SLC34A2 O95436 Non-Small Cell Lung cancer, Ovarian Cancer SLC39A6 Q13433 Breast Cancer CanAg (a glycoform of P15941 Breast Cancer MUC1) Mucin 16 (CA125) Q8WXI7 Epithelial Ovarian Cancer, Breast Cancer CAIX Q16790 Renal Cell Carcinoma RAAG12 N-linked carbohydrate Adenocarcinoma epitope Sialyl LewisA carbohydrate epitope Gastrointestinal cancers Lewis Y (Le(y)) antigen carbohydrate epitope Gastrointestinal cancers Autoimmune disease/Inflammation CD19 see above autoimmune disease CD20 see above Rheumatoid Arthritis CD70 (CD27L) see above Autoimmune Diseases, Cancer, Inflammatory Diseases CD79b see above Systemic lupus erythematosus IL-5R.alpha. Q01344 Asthma, Chronic obstructive pulmonary disease IL-23R Q9NPF7 Inflammatory Diseases, Autoimmune Diseases, Cancer TF (Tissue Factor) see above Acute Lung Injury, Inflammatory Diseases TIM-3 see above Autoimmunity, Inflammation Viral infections- human targets TSG101 Q99816 HIV, Herpes, Influenza, Ebola WNV E Q91KZ4 West Nile Virus CD81 P60033 HCV (entry) CD4 P01730 HIV CXCR4 P61073 HIV CCR5 P51681 HIV Integrin .alpha.L P20701 HIV

Scaffolds

[0114] In some embodiments, the antigen-binding constructs described herein comprise a scaffold. A scaffold may be a peptide, polypeptide, polymer, nanoparticle or other chemical entity. In embodiments where the scaffold is an Fc or dimeric Fc, the antigen-binding polypeptide construct(s) of the antigen-binding construct may be linked to either the N- or C-terminus of the scaffold. A dimeric Fc can be homodimeric or heterodimeric.

[0115] In embodiments where the scaffold is a peptide or polypeptide, the antigen-binding construct or antigen-binding polypeptide construct may be linked to the scaffold by genetic fusion with or without polypeptide linkers. In other embodiments, where the scaffold is a polymer or nanoparticle, the antigen-binding construct may be linked to the scaffold by chemical conjugation. In some embodiments, the scaffold is an albumin polypeptide or split albumin polypeptide. The use of split albumin polypeptides as scaffolds for antigen-binding polypeptide constructs is fully described in PCT/CA2012/050131, PCT/US2013/050408 and PCT/US2013/050411 all of which are hereby incorporated by reference in their entirety.

Fc of Antigen-Binding Constructs.

[0116] Fc polypeptides make excellent scaffolds for antigen-binding polypeptide constructs. Certain antigen-binding constructs described herein comprise an Fc, e.g., a dimeric Fc. In some embodiments, the Fc is a a heterodimeric Fc comprising first and second Fc polypeptides each comprising a modified CH3 sequence, wherein each modified CH3 sequence comprises asymmetric amino acid modifications that promote the formation of a heterodimeric Fc and the dimerized CH3 domains have a melting temperature (Tm) of about 68.degree. C. or higher, and wherein the first Fc polypeptide is linked to the first antigen-binding polypeptide construct, with a first hinge linker, and the second Fc polypeptide is linked to the second antigen-binding polypeptide construct with a second hinge linker.

[0117] The term "Fc domain" or "Fc region" herein is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. Unless otherwise specified herein, numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also called the EU index, as described in Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991. An "Fc polypeptide" of a dimeric Fc as used herein refers to one of the two polypeptides forming the dimeric Fc domain, i.e. a polypeptide comprising C-terminal constant regions of an immunoglobulin heavy chain, capable of stable self-association. For example, an Fc polypeptide of a dimeric IgG Fc comprises an IgG CH2 and an IgG CH3 constant domain sequence.

[0118] An Fc domain comprises either a CH3 domain or a CH3 and a CH2 domain. The CH3 domain comprises two CH3 sequences, one from each of the two Fc polypeptides of the dimeric Fc. The CH2 domain comprises two CH2 sequences, one from each of the two Fc polypeptides of the dimeric Fc.

[0119] In some aspects, the Fc comprises at least one or two CH3 sequences. In some aspects, the Fc is coupled, with or without one or more linkers, to a first antigen-binding construct and/or a second antigen-binding construct. In some aspects, the Fc is a human Fc. In some aspects, the Fc is a human IgG or IgG1 Fc. In some aspects, the Fc is a heterodimeric Fc. In some aspects, the Fc comprises at least one or two CH2 sequences.

[0120] In some aspects, the Fc comprises one or more modifications in at least one of the CH3 sequences. In some aspects, the Fc comprises one or more modifications in at least one of the CH2 sequences. In some aspects, an Fc is a single polypeptide. In some aspects, an Fc is multiple peptides, e.g., two polypeptides.

[0121] In some aspects, the Fc is an Fc described in patent applications PCT/CA2011/001238, filed Nov. 4, 2011 or PCT/CA2012/050780, filed Nov. 2, 2012, the entire disclosure of each of which is hereby incorporated by reference in its entirety for all purposes.

[0122] Modified CH3 Domains

[0123] In some aspects, the antigen-binding construct described herein comprises a heterodimeric Fc comprising a modified CH3 domain that has been asymmetrically modified. The heterodimeric Fc can comprise two heavy chain constant domain polypeptides: a first Fc polypeptide and a second Fc polypeptide, which can be used interchangeably provided that Fc comprises one first Fc polypeptide and one second Fc polypeptide. Generally, the first Fc polypeptide comprises a first CH3 sequence and the second Fc polypeptide comprises a second CH3 sequence.

[0124] Two CH3 sequences that comprise one or more amino acid modifications introduced in an asymmetric fashion generally results in a heterodimeric Fc, rather than a homodimer, when the two CH3 sequences dimerize. As used herein, "asymmetric amino acid modifications" refers to any modification where an amino acid at a specific position on a first CH3 sequence is different from the amino acid on a second CH3 sequence at the same position, and the first and second CH3 sequence preferentially pair to form a heterodimer, rather than a homodimer. This heterodimerization can be a result of modification of only one of the two amino acids at the same respective amino acid position on each sequence; or modification of both amino acids on each sequence at the same respective position on each of the first and second CH3 sequences. The first and second CH3 sequence of a heterodimeric Fc can comprise one or more than one asymmetric amino acid modification.

[0125] Table C provides the amino acid sequence of the human IgG1 Fc sequence, corresponding to amino acids 231 to 447 of the full-length human IgG1 heavy chain. Amino acids 231-238 are also referred to as the lower hinge. The CH3 sequence comprises amino acid 341-447 of the full-length human IgG1 heavy chain.

[0126] Typically an Fc can include two contiguous heavy chain sequences (A and B) that are capable of dimerizing. With respect to the antigen binding constructs described herein, in some embodiments the first scFv is linked to chain A of the heterodimeric Fc and the second scFv is linked to chain B of the heterodimeric Fc. in some embodiments the second scFv is linked to chain A of the heterodimeric Fc and the first scFv is linked to chain B of the heterodimeric Fc.

[0127] In some aspects, one or both sequences of an Fc include one or more mutations or modifications at the following locations: L351, F405, Y407, T366, K392, T394, T350, 5400, and/or N390, using EU numbering. In some aspects, an Fc includes a mutant sequence shown in Table X. In some aspects, an Fc includes the mutations of Variant 1 A-B. In some aspects, an Fc includes the mutations of Variant 2 A-B. In some aspects, an Fc includes the mutations of Variant 3 A-B. In some aspects, an Fc includes the mutations of Variant 4 A-B. In some aspects, an Fc includes the mutations of Variant 5 A-B.

TABLE-US-00006 TABLE C IgG1 Fc sequence and variants Human IgG1 Fc APELLGGPSVFLFPPKPKDTLMIS sequence 231-447 RTPEVTCVVVDVSHEDPEVKFNWY (EU-numbering) VDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNK ALPAPIEKTISKAKGQPREPQVYT LPPSRDELTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPG K (SEQ ID NO: 361) Variant IgG1 Fc sequence (231-447) Chain Mutations 1 A L351Y_F405A_Y407V 1 B T366L_K392M_T394W 2 A L351Y_F405A_Y407V 2 B T366L_K392L_T394W 3 A T350V_L351Y_F405A_Y407V 3 B T350V_T366L_K392L_T394W 4 A T350V_L351Y_F405A_Y407V 4 B T350V_T366L_K392M_T394W 5 A T350V_L351Y_S400E_F405A_Y407V 5 B T350V_T366L_N390R_K392M_T394W

[0128] The first and second CH3 sequences can comprise amino acid mutations as described herein, with reference to amino acids 231 to 447 of the full-length human IgG1 heavy chain. In one embodiment, the heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having amino acid modifications at positions F405 and Y407, and a second CH3 sequence having amino acid modifications at position T394. In one embodiment, the heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having one or more amino acid modifications selected from L351Y, F405A, and Y407V, and the second CH3 sequence having one or more amino acid modifications selected from T366L, T366I, K392L, K392M, and T394W.

[0129] In one embodiment, a heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having amino acid modifications at positions L351, F405 and Y407, and a second CH3 sequence having amino acid modifications at positions T366, K392, and T394, and one of the first or second CH3 sequences further comprising amino acid modifications at position Q347, and the other CH3 sequence further comprising amino acid modification at position K360. In another embodiment, a heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having amino acid modifications at positions L351, F405 and Y407, and a second CH3 sequence having amino acid modifications at position T366, K392, and T394, one of the first or second CH3 sequences further comprising amino acid modifications at position Q347, and the other CH3 sequence further comprising amino acid modification at position K360, and one or both of said CH3 sequences further comprise the amino acid modification T350V.

[0130] In one embodiment, a heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having amino acid modifications at positions L351, F405 and Y407, and a second CH3 sequence having amino acid modifications at positions T366, K392, and T394 and one of said first and second CH3 sequences further comprising amino acid modification of D399R or D399K and the other CH3 sequence comprising one or more of T411E, T411D, K409E, K409D, K392E and K392D. In another embodiment, a heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having amino acid modifications at positions L351, F405 and Y407, and a second CH3 sequence having amino acid modifications at positions T366, K392, and T394, one of said first and second CH3 sequences further comprises amino acid modification of D399R or D399K and the other CH3 sequence comprising one or more of T411E, T411D, K409E, K409D, K392E and K392D, and one or both of said CH3 sequences further comprise the amino acid modification T350V.

[0131] In one embodiment, a heterodimeric Fc comprises a modified CH3 domain with a first CH3 sequence having amino acid modifications at positions L351, F405 and Y407, and a second CH3 sequence having amino acid modifications at positions T366, K392, and T394, wherein one or both of said CH3 sequences further comprise the amino acid modification of T350V.

[0132] In one embodiment, a heterodimeric Fc comprises a modified CH3 domain comprising the following amino acid modifications, where "A" represents the amino acid modifications to the first CH3 sequence, and "B" represents the amino acid modifications to the second CH3 sequence: A:L351Y_F405A_Y407V, B:T366L_K392M_T394W, A:L351Y_F405A_Y407V, B:T366L_K392L_T394W, A:T350V_L351Y_F405A_Y407V, B:T350V_T366L_K392L_T394W, A:T350V_L351Y_F405A_Y407V, B:T350V_T366L_K392M_T394W, A:T350V_L351Y_S400E_F405A_Y407V, and/or B:T350V_T366L_N390R_K392M_T394W.

[0133] The one or more asymmetric amino acid modifications can promote the formation of a heterodimeric Fc in which the heterodimeric CH3 domain has a stability that is comparable to a wild-type homodimeric CH3 domain. In an embodiment, the one or more asymmetric amino acid modifications promote the formation of a heterodimeric Fc domain in which the heterodimeric Fc domain has a stability that is comparable to a wild-type homodimeric Fc domain. In an embodiment, the one or more asymmetric amino acid modifications promote the formation of a heterodimeric Fc domain in which the heterodimeric Fc domain has a stability observed via the melting temperature (Tm) in a differential scanning calorimetry study, and where the melting temperature is within 4.degree. C. of that observed for the corresponding symmetric wild-type homodimeric Fc domain. In some aspects, the Fc comprises one or more modifications in at least one of the CH3 sequences that promote the formation of a heterodimeric Fc with stability comparable to a wild-type homodimeric Fc.

[0134] In one embodiment, the stability of the CH3 domain can be assessed by measuring the melting temperature of the CH3 domain, for example by differential scanning calorimetry (DSC). Thus, in a further embodiment, the CH3 domain has a melting temperature of about 68.degree. C. or higher. In another embodiment, the CH3 domain has a melting temperature of about 70.degree. C. or higher. In another embodiment, the CH3 domain has a melting temperature of about 72.degree. C. or higher. In another embodiment, the CH3 domain has a melting temperature of about 73.degree. C. or higher. In another embodiment, the CH3 domain has a melting temperature of about 75.degree. C. or higher. In another embodiment, the CH3 domain has a melting temperature of about 78.degree. C. or higher. In some aspects, the dimerized CH3 sequences have a melting temperature (Tm) of about 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 77.5, 78, 79, 80, 81, 82, 83, 84, or 85.degree. C. or higher.

[0135] In some embodiments, a heterodimeric Fc comprising modified CH3 sequences can be formed with a purity of at least about 75% as compared to homodimeric Fc in the expressed product. In another embodiment, the heterodimeric Fc is formed with a purity greater than about 80%. In another embodiment, the heterodimeric Fc is formed with a purity greater than about 85%. In another embodiment, the heterodimeric Fc is formed with a purity greater than about 90%. In another embodiment, the heterodimeric Fc is formed with a purity greater than about 95%. In another embodiment, the heterodimeric Fc is formed with a purity greater than about 97%. In some aspects, the Fc is a heterodimer formed with a purity greater than about 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% when expressed. In some aspects, the Fc is a heterodimer formed with a purity greater than about 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% when expressed via a single cell.

[0136] Additional methods for modifying monomeric Fc polypeptides to promote heterodimeric Fc formation are known in the art. For example, see International Patent Publication No. WO 96/027011 (knobs into holes), in Gunasekaran et al. (Gunasekaran K. et al. (2010) J Biot Chem. 285, 19637-46, electrostatic design to achieve selective heterodimerization), in Davis et al. (Davis, J H. et al. (2010) Prot Eng Des Set; 23(4): 195-202, strand exchange engineered domain (SEED) technology), and in Labrijn et al [Efficient generation of stable bi-specific IgG1 by controlled Fab-arm exchange. Labrijn A F, Meesters J I, de Goeij B E, van den Bremer E T, Neijssen J, van Kampen M D, Strumane K, Verploegen S, Kundu A, Cramer M J, van Berkel P H, van de Winkel J G, Schuurman J, Parren P W. Proc Natl Acad Sci USA. 2013 Mar. 26; 110(13):5145-50.

[0137] CH2 Domains

[0138] As indicated above, in some embodiments, the Fc of the antigen-binding construct comprises a CH2 domain in addition to a CH3 domain. As an example, the amino acid sequence of the CH2 domain of an IgG1 Fc is identified as amino acids 239-340 of the sequence shown in Table A. The CH2 domain of the Fc binds to Fc receptors and complement and is thus involved in mediating effector cell functions.

[0139] The terms "Fc receptor" and "FcR" are used to describe a receptor that binds to the Fc region of an antibody, and includes Fc gamma receptors (Fc.gamma.Rs) and the neonatal receptor FcRn.

[0140] Generally, an Fc.gamma.R is one which binds an IgG antibody (a gamma receptor) and includes receptors of the Fc.gamma.RI, Fc.gamma.RII, and Fc.gamma.RIII subclasses in humans, including allelic variants and alternatively spliced forms of these receptors. Fc.gamma.RII receptors include Fc.gamma.RIIA (an "activating receptor") and Fc.gamma.RIIB (an "inhibiting receptor"), which have similar amino acid sequences that differ primarily in the cytoplasmic domains thereof. Immunoglobulins of other isotypes can also be bound by certain FcRs (see, e.g., Janeway et al., Immuno Biology: the immune system in health and disease, (Elsevier Science Ltd., NY) (4th ed., 1999)). Activating receptor Fc.gamma.RIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor Fc.gamma.RIIB contains an immunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain (reviewed in Daeron, Annu. Rev. Immunol. 15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126:330-41 (1995). Other Fc.gamma.Rs, including those to be identified in the future, are encompassed by the term "FcR" herein. An Fc.gamma.R are also found in other organisms, including but not limited to mice, rats, rabbits, and monkeys. Mouse Fc.gamma.Rs include but are not limited to Fc.gamma.RI (CD64), Fc.gamma.RII (CD32), Fc.gamma.RIII (CD 16), and Fc.gamma.RIII-2 (CD 16-2). Fc.gamma.Rs are expressed by effector cells such as NK cells or B cells.

[0141] Complement activation requires binding of the complement protein C1q to antigen-antibody complexes. Residues in the CH2 domain of the Fc are involved in the interaction between C1q and the Fc.

[0142] Some of the antigen-binding constructs described herein are able to bind FcRn. As is known in the art, binding to FcRn recycles endocytosed antibody from the endosome back to the bloodstream (Raghavan et al., 1996, Annu Rev Cell Dev Biol 12:181-220; Ghetie et al., 2000, Annu Rev Immunol 18:739-766). This process, coupled with preclusion of kidney filtration due to the large size of the full-length molecule, results in favorable antibody serum half-lives ranging from one to three weeks. Binding of Fc to FcRn also plays a key role in antibody transport. FcRn is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J. Immunol. 117:587 (1976); and Kim et al., J. Immunol. 24:249 (1994)). Binding of the FcRn to IgG involves residues in the CH2 and CH3 domains of the Fc.

[0143] Modifications in the CH2 domain can affect the binding of FcRs to the Fc. As indicated above, the CH2 domain of the Fc comprises two CH2 sequences, one on each of the two Fc polypeptides of the dimeric Fc. Typically, the modifications to the CH2 domain are symmetric and are thus the same on both CH2 sequences of the Fc polypeptides. However, asymmetric mutations are also possible in the presence of mutations on the CH3 domain that enhance heterodimerization. In one embodiment, the CH2 domain comprises modifications to reduce Fc.gamma.R or C1q binding and/or effector function.

Modifications to Reduce Effector Function:

[0144] Fc modifications reducing Fc.gamma.R and/or complement binding and/or effector function are known in the art. Recent publications describe strategies that have been used to engineer antibodies with reduced or silenced effector activity (see Strohl, W R (2009), Curr Opin Biotech 20:685-691, and Strohl, W R and Strohl L M, "Antibody Fc engineering for optimal antibody performance" In Therapeutic Antibody Engineering, Cambridge: Woodhead Publishing (2012), pp 225-249). These strategies include reduction of effector function through modification of glycosylation, use of IgG2/IgG4 scaffolds, or the introduction of mutations in the hinge or CH2 regions of the Fc. For example, US Patent Publication No. 2011/0212087 (Strohl), International Patent Publication No. WO 2006/105338 (Xencor), US Patent Publication No. 2012/0225058 (Xencor), US Patent Publication No. 2012/0251531 (Genentech), and Strop et al ((2012) J. Mol. Biol. 420: 204-219) describe specific modifications to reduce Fc.gamma.R or complement binding to the Fc.

[0145] Specific, non-limiting examples of known symmetric amino acid modifications to reduce Fc.gamma.R or complement binding to the Fc include those identified in the following table:

TABLE-US-00007 TABLE D modifications to reduce Fc.gamma.R or complement binding to the Fc Company Mutations GSK N297A Ortho Biotech L234A/L235A Protein Design labs IGG2 V234A/G237A Wellcome Labs IGG4 L235A/G237A/E318A GSK IGG4 S228P/L236E Alexion IGG2/IgG4 combination Merck IGG2 H268Q/V309L/A330S/A331S Bristol-Myers C220S/C226S/C229S/P238S Seattle Genetics C226S/C229S/E3233P/L235V/L235A Amgen E. coli production, non glycosylated Medimune L234F/L235E/P331S Trubion Hinge mutant, possibly C226S/P230S

[0146] In one embodiment, the Fc comprises at least one amino acid modification identified in the above table. In another embodiment the Fc comprises amino acid modification of at least one of L234, L235, or D265. In another embodiment, the Fc comprises amino acid modification at L234, L235 and D265. In another embodiment, the Fc comprises the amino acid modifications L234A, L235A and D265S.

[0147] In some embodiments the Fc comprises one or more asymmetric amino acid modifications in the lower hinge region of the Fc as described in International Patent Application No. PCT/CA2014/050507. Examples of such asymmetric amino acid modifications that reduce Fc.gamma.R binding are shown in Table E:

TABLE-US-00008 TABLE E Asymmetric mutations that reduce Fc.gamma.R binding Chain A Chain B L234D/L235E L234K/L235K E233A/L234D/L235E E233A/L234R/L235R L234D/L235E E233K/L234R/L235R E233A/L234K/L235A E233K/L234A/L235K

Hinge Linkers

[0148] In the antigen-binding constructs described herein, the first Fc polypeptide is linked to the first antigen-binding polypeptide construct with a first hinge linker, and the second Fc polypeptide is linked to the second antigen-binding polypeptide construct with a second hinge linker. Examples of hinge linker sequences are well-known to one of skill in the art and can be used in the antigen-binding constructs described herein. Alternatively, modified versions of known hinge linkers can be used.

[0149] The hinge linker polypeptides are selected such that they maintain or optimize the functional activity of the antigen-binding construct. Suitable linker polypeptides include IgG hinge regions such as, for example those from IgG.sub.1, IgG.sub.2, or IgG.sub.4, including the upper hinge sequences and core hinge sequences. The amino acid residues corresponding to the upper and core hinge sequences vary depending on the IgG type, as is known in the art and one of skill in the art would readily be able to identify such sequences for a given IgG type. Modified versions of these exemplary linkers can also be used. For example, modifications to improve the stability of the IgG.sub.4 hinge are known in the art (see for example, Labrijn et al. (2009) Nature Biotechnology 27, 767-771). Examples of hinge linker sequences are found in the following Table. In some embodiments, the drug-conjugated antigen-binding constructs described herein have modifications to the hinge region to modify or optimize potency of the construct.

TABLE-US-00009 TABLE F Hinge linker polypeptide sequences (SEQ ID NOS:) SEQ ID NO: 995 IgG1 EPKSCDKTHTCPPCP 996 IgG1 GAGCCCAAGAGCTGTGATAAGACCC ACACCTGCCCTCCCTGTCCA 997 v1661 AAEPKSSDKTHTCPPCP 998 v1661 GCAGCCGAACCCAAATCCTCTGATA AGACCCACACATGCCCTCCATGTCC A 999 Hinge-1 EPKSSDKTHTCPPCP 1000 Hinge-1 GAGCCTAAAAGCTCCGACAAGACCC ACACATGCCCACCTTGTCCG 1001 Hinge-2 DKTHTCPPCP 1002 Hinge-2 GACAAGACCCACACATGCCCACCTT GTCCG 1003 Hinge-3 GTCPPCP 1004 Hinge-3 GGCACATGCCCTCCATGTCCA

Dissociation Constant (K.sub.D) and Maximal Binding (Bmax)

[0150] In some embodiments, an antigen-binding construct is described by functional characteristics including but not limited to a dissociation constant and a maximal binding.

[0151] The term "dissociation constant (K.sub.D)" as used herein, is intended to refer to the equilibrium dissociation constant of a particular ligand-protein interaction. As used herein, ligand-protein interactions refer to, but are not limited to protein-protein interactions or antibody-antigen interactions. The K.sub.D measures the propensity of two proteins (e.g. AB) to dissociate reversibly into smaller components (A+B), and is define as the ratio of the rate of dissociation, also called the "off-rate (k.sub.off)", to the association rate, or "on-rate (k.sub.on)". Thus, K.sub.D equals k.sub.off/k.sub.on and is expressed as a molar concentration (M). It follows that the smaller the K.sub.D, the stronger the affinity of binding. Therefore, a K.sub.D of 1 mM indicates weak binding affinity compared to a K.sub.D of 1 nM. K.sub.D values for antigen-binding constructs can be determined using methods well established in the art. One method for determining the K.sub.D of an antigen-binding construct is by using surface plasmon resonance (SPR), typically using a biosensor system such as a Biacore.RTM. system. Isothermal titration calorimetry (ITC) is another method that can be used to determine.

[0152] The term "Bmax", or maximal binding, refers to the maximum antigen-binding construct binding level on the cells at saturating concentrations of antigen-binding construct. This parameter can be reported in the arbitrary unit MFI for relative comparison, or converted into an absolute value corresponding to the number of antigen-binding constructs bound to the cell with the use of a standard curve.

[0153] The binding characteristics of an antigen-binding construct can be determined by various techniques. One of which is the measurement of binding to target cells expressing the antigen by flow cytometry (FACS, Fluorescence-activated cell sorting). Typically, in such an experiment, the target cells expressing the antigen of interest are incubated with antigen-binding constructs at different concentrations, washed, incubated with a secondary agent for detecting the antigen-binding construct, washed, and analyzed in the flow cytometer to measure the median fluorescent intensity (MFI) representing the strength of detection signal on the cells, which in turn is related to the number of antigen-binding constructs bound to the cells. The antigen-binding construct concentration vs. MFI data is then fitted into a saturation binding equation to yield two key binding parameters, Bmax and apparent K.sub.D.

[0154] Apparent K.sub.D, or apparent equilibrium dissociation constant, represents the antigen-binding construct concentration at which half maximal cell binding is observed. Evidently, the smaller the K.sub.D value, the smaller antigen-binding construct concentration is required to reach maximum cell binding and thus the higher is the affinity of the antigen-binding construct. The apparent K.sub.D is dependent on the conditions of the cell binding experiment, such as different receptor levels expressed on the cells and incubation conditions, and thus the apparent K.sub.D is generally different from the K.sub.D values determined from cell-free molecular experiments such as SPR and ITC. However, there is generally good agreement between the different methods.

[0155] In some embodiments of a drug-conjugated antigen-binding construct described herein, one antigen-binding polypeptide construct has a higher affinity for its cognate antigen than the other. In most embodiments of a drug-conjugated antigen-binding construct, the first antigen-binding polypeptide construct has a lower affinity for CD3 than the second antigen-binding polypeptide construct has for the target antigen. In some embodiments, the construct has at least 2, at least 5, at least 10, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90 or at least 100-fold lower affinity for the CD3 antigen than for the target antigen, as measured by SPR; and/or has an an affinity of less than 10 nM for target cells bearing the target antigen and an affinity in the range of 10 nM-500 nM for T cells as measured by FACS.

[0156] In many embodiments, the affinity for CD3 will be lower than the affinity for the target antigen. In one embodiment of a CD3-CD19 drug-conjugated antigen-binding construct, the affinity for CD3 is lower than the affinity for CD19. In further embodiments, the affinity for CD3 is at least 2, 5, 10, 15 or 20-fold lower than the affinity for CD19. In one specific embodiment, the affinity of a CD3-CD19 drug-conjugated antigen-binding construct is 2 nM for CD19 and 30 nM for CD3. Affinities may be determined by SPR. In some embodiments the affinity of the second antigen-binding polypeptide construct for CD19 antigen expressed on a B cell is in the range of about 0.5-1, 1-3, 3-5, 5-7, 7-9, 9-11, 11-13, 13-15, 15-17, 17-19 or 19-21 nM, and the affinity of the first antigen-binding polypeptide construct for CD3 expressed on a T cell is in the range of about 5-10, 10-15, 15-20, 20-15, 25-30, 30-35, 35-40, 40-50, 50-55, 55-60, 60-70, 70-80, 80-90 or 90-100 nm, as determined by FACS analysis.

Methods of Preparation of Antigen-Binding Constructs

[0157] Antigen-binding constructs described herein may be produced using recombinant methods and compositions, e.g., as described in U.S. Pat. No. 4,816,567.

[0158] In one embodiment, an isolated nucleic acid encoding an antigen-binding construct described herein is provided. Such nucleic acid may encode an amino acid sequence comprising the VL and/or an amino acid sequence comprising the VH of the antigen-binding construct (e.g., the light and/or heavy chains of the antigen-binding construct). In a further embodiment, one or more vectors (e.g., expression vectors) comprising such nucleic acid are provided. In one embodiment, the nucleic acid is provided in a multicistronic vector. In a further embodiment, a host cell comprising such nucleic acid is provided. In one such embodiment, a host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antigen-binding construct and an amino acid sequence comprising the VH of the antigen-binding polypeptide construct, or (2) a first vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antigen-binding polypeptide construct and a second vector comprising a nucleic acid that encodes an amino acid sequence comprising the VH of the antigen-binding polypeptide construct. In one embodiment, the host cell is eukaryotic, e.g. a Chinese Hamster Ovary (CHO) cell, or human embryonic kidney (HEK) cell, or lymphoid cell (e.g., Y0, NS0, Sp20 cell). In one embodiment, a method of making an antigen-binding construct is provided, wherein the method comprises culturing a host cell comprising nucleic acid encoding the antigen-binding construct, as provided above, under conditions suitable for expression of the antigen-binding construct, and optionally recovering the antigen-binding construct from the host cell (or host cell culture medium).

[0159] For recombinant production of the antigen-binding construct, a nucleic acid encoding an antigen-binding construct, e.g., as described above, is isolated and inserted into one or more vectors for further cloning and/or expression in a host cell. Such nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antigen-binding construct).

[0160] Suitable host cells for cloning or expression of antigen-binding construct-encoding vectors include prokaryotic or eukaryotic cells described herein.

[0161] A "recombinant host cell" or "host cell" refers to a cell that includes an exogenous polynucleotide, regardless of the method used for insertion, for example, direct uptake, transduction, f-mating, or other methods known in the art to create recombinant host cells. The exogenous polynucleotide may be maintained as a nonintegrated vector, for example, a plasmid, or alternatively, may be integrated into the host genome.

[0162] As used herein, the term "eukaryote" refers to organisms belonging to the phylogenetic domain Eucarya such as animals (including but not limited to, mammals, insects, reptiles, birds, etc.), ciliates, plants (including but not limited to, monocots, dicots, algae, etc.), fungi, yeasts, flagellates, microsporidia, protists, etc.

[0163] As used herein, the term "prokaryote" refers to prokaryotic organisms. For example, a non-eukaryotic organism can belong to the Eubacteria (including but not limited to, Escherichia coli, Thermus thermophilus, Bacillus stearothermophilus, Pseudomonas fluorescens, Pseudomonas aeruginosa, Pseudomonas putida, etc.) phylogenetic domain, or the Archaea (including but not limited to, Methanococcus jannaschii, Methanobacterium thermoautotrophicum, Halobacterium such as Haloferax volcanii and Halobacterium species NRC-1, Archaeoglobus fulgidus, Pyrococcus furiosus, Pyrococcus horikoshii, Aeuropyrum pernix, etc.) phylogenetic domain.

[0164] For example, antigen-binding constructs may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed. For expression of antigen-binding construct fragments and polypeptides in bacteria, see, e.g., U.S. Pat. Nos. 5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B. K. C. Lo, ed., Humana Press, Totowa, N.J., 2003), pp. 245-254, describing expression of antibody fragments in E. coli.) After expression, the antigen-binding construct may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.

[0165] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antigen-binding construct-encoding vectors, including fungi and yeast strains whose glycosylation pathways have been "humanized," resulting in the production of an antigen-binding construct with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., Nat. Biotech. 24:210-215 (2006).

[0166] Suitable host cells for the expression of glycosylated antigen-binding constructs are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

[0167] Plant cell cultures can also be utilized as hosts. See, e.g., U.S. Pat. Nos. 5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIES.TM. technology for producing antigen-binding constructs in transgenic plants).

[0168] Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol. 36:59 (1977)); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR.sup.- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0 and Sp2/0. For a review of certain mammalian host cell lines suitable for antigen-binding construct production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B. K. C. Lo, ed., Humana Press, Totowa, N.J.), pp. 255-268 (2003).

[0169] In one embodiment, the antigen-binding constructs described herein are produced in stable mammalian cells, by a method comprising: transfecting at least one stable mammalian cell with: nucleic acid encoding the antigen-binding construct, in a predetermined ratio; and expressing the nucleic acid in the at least one mammalian cell. In some embodiments, the predetermined ratio of nucleic acid is determined in transient transfection experiments to determine the relative ratio of input nucleic acids that results in the highest percentage of the antigen-binding construct in the expressed product.

[0170] If required, the antigen-binding constructs can be purified or isolated after expression. Proteins may be isolated or purified in a variety of ways known to those skilled in the art. Standard purification methods include chromatographic techniques, including ion exchange, hydrophobic interaction, affinity, sizing or gel filtration, and reversed-phase, carried out at atmospheric pressure or at high pressure using systems such as FPLC and HPLC. Purification methods also include electrophoretic, immunological, precipitation, dialysis, and chromatofocusing techniques. Ultrafiltration and diafiltration techniques, in conjunction with protein concentration, are also useful. As is well known in the art, a variety of natural proteins bind Fc and antibodies, and these proteins can find use in the present invention for purification of antigen-binding constructs. For example, the bacterial proteins A and G bind to the Fc region. Likewise, the bacterial protein L binds to the Fab region of some antibodies. Purification can often be enabled by a particular fusion partner. For example, antibodies may be purified using glutathione resin if a GST fusion is employed, Ni.sup.+2 affinity chromatography if a His-tag is employed, or immobilized anti-flag antibody if a flag-tag is used. For general guidance in suitable purification techniques, see, e.g. incorporated entirely by reference Protein Purification: Principles and Practice, 3.sup.rd Ed., Scopes, Springer-Verlag, NY, 1994, incorporated entirely by reference. The degree of purification necessary will vary depending on the use of the antigen-binding constructs. In some instances no purification is necessary.

[0171] In certain embodiments the antigen-binding constructs are purified using Anion Exchange Chromatography including, but not limited to, chromatography on Q-sepharose, DEAE sepharose, poros HQ, poros DEAF, Toyopearl Q, Toyopearl QAE, Toyopearl DEAE, Resource/Source Q and DEAE, Fractogel Q and DEAE columns.

[0172] In specific embodiments the proteins described herein are purified using Cation Exchange Chromatography including, but not limited to, SP-sepharose, CM sepharose, poros HS, poros CM, Toyopearl SP, Toyopearl CM, Resource/Source S and CM, Fractogel S and CM columns and their equivalents and comparables.

[0173] In addition, antigen-binding constructs described herein can be chemically synthesized using techniques known in the art (e.g., see Creighton, 1983, Proteins: Structures and Molecular Principles, W. H. Freeman & Co., N.Y and Hunkapiller et al., Nature, 310:105-111 (1984)). For example, a polypeptide corresponding to a fragment of a polypeptide can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the polypeptide sequence. Non-classical amino acids include, but are not limited to, to the D-isomers of the common amino acids, 2,4diaminobutyric acid, alpha-amino isobutyric acid, 4aminobutyric acid, Abu, 2-amino butyric acid, g-Abu, e-Ahx, 6amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, .right brkt-bot.-alanine, fluoro-amino acids, designer amino acids such as .right brkt-bot.-methyl amino acids, C .right brkt-bot.-methyl amino acids, N .right brkt-bot.-methyl amino acids, and amino acid analogs in general. Furthermore, the amino acid can be D (dextrorotary) or L (levorotary).

[0174] In some embodiments, the antigen-binding constructs described herein are substantially purified. The term "substantially purified" refers to a construct described herein, or variant thereof that may be substantially or essentially free of components that normally accompany or interact with the protein as found in its naturally occurring environment, i.e. a native cell, or host cell in the case of recombinantly produced antigen-binding construct that in certain embodiments, is substantially free of cellular material includes preparations of protein having less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% (by dry weight) of contaminating protein. When the antigen-binding construct or variant thereof is recombinantly produced by the host cells, the protein in certain embodiments is present at about 30%, about 25%, about 20%, about 15%, about 10%, about 5%, about 4%, about 3%, about 2%, or about 1% or less of the dry weight of the cells. When the antigen-binding construct or variant thereof is recombinantly produced by the host cells, the protein, in certain embodiments, is present in the culture medium at about 5 g/L, about 4 g/L, about 3 g/L, about 2 g/L, about 1 g/L, about 750 mg/L, about 500 mg/L, about 250 mg/L, about 100 mg/L, about 50 mg/L, about 10 mg/L, or about 1 mg/L or less of the dry weight of the cells. In certain embodiments, a "substantially purified" antigen-binding construct produced by the methods described herein, has a purity level of at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, specifically, a purity level of at least about 75%, 80%, 85%, and more specifically, a purity level of at least about 90%, a purity level of at least about 95%, a purity level of at least about 99% or greater as determined by appropriate methods such as SDS/PAGE analysis, RP-HPLC, SEC, and capillary electrophoresis.

[0175] Post-Translational Modifications:

[0176] In certain embodiments antigen-binding constructs described herein are differentially modified during or after translation.

[0177] The term "modified," as used herein refers to any changes made to a given polypeptide, such as changes to the length of the polypeptide, the amino acid sequence, chemical structure, co-translational modification, or post-translational modification of a polypeptide. The form "(modified)" term means that the polypeptides being discussed are optionally modified, that is, the polypeptides under discussion can be modified or unmodified.

[0178] The term "post-translationally modified" refers to any modification of a natural or non-natural amino acid that occurs to such an amino acid after it has been incorporated into a polypeptide chain. The term encompasses, by way of example only, co-translational in vivo modifications, co-translational in vitro modifications (such as in a cell-free translation system), post-translational in vivo modifications, and post-translational in vitro modifications.

[0179] In some embodiments, the modification is at least one of: glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage and linkage to an antibody molecule or antigen-binding construct or other cellular ligand. In some embodiments, the antigen-binding construct is chemically modified by known techniques, including but not limited, to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease, NaBH.sub.4; acetylation, formylation, oxidation, reduction; and metabolic synthesis in the presence of tunicamycin.

[0180] Additional post-translational modifications of antigen-binding constructs described herein include, for example, N-linked or O-linked carbohydrate chains, processing of N-terminal or C-terminal ends), attachment of chemical moieties to the amino acid backbone, chemical modifications of N-linked or O-linked carbohydrate chains, and addition or deletion of an N-terminal methionine residue as a result of procaryotic host cell expression. The antigen-binding constructs described herein are modified with a detectable label, such as an enzymatic, fluorescent, isotopic or affinity label to allow for detection and isolation of the protein. In certain embodiments, examples of suitable enzyme labels include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; and examples of suitable radioactive material include iodine, carbon, sulfur, tritium, indium, technetium, thallium, gallium, palladium, molybdenum, xenon, fluorine.

[0181] In some embodiments, antigen-binding constructs described herein are attached to macrocyclic chelators that associate with radiometal ions.

[0182] In some embodiments, the antigen-binding constructs described herein are modified by either natural processes, such as post-translational processing, or by chemical modification techniques which are well known in the art. In certain embodiments, the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. In certain embodiments, polypeptides from antigen-binding constructs described herein are branched, for example, as a result of ubiquitination, and in some embodiments are cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides are a result from posttranslation natural processes or made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, for instance, PROTEINS--STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993); POST-TRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, pgs. 1-12 (1983); Seifter et al., Meth. Enzymol. 182:626-646 (1990); Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62 (1992)).

[0183] In certain embodiments, antigen-binding constructs described herein are attached to solid supports, which are particularly useful for immunoassays or purification of polypeptides that are bound by, that bind to, or associate with proteins described herein. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene.

Functional Activity of Drug-Conjugated Antigen-Binding Constructs and Assays to Measure Function

[0184] The antigen-binding constructs described herein can be assayed for functional activity (e.g., biological activity) using or routinely modifying assays known in the art, as well as assays described herein.

[0185] Methods of testing the biological activity of the antigen-binding constructs and drug-conjugated antigen-binding constructs described herein can be measured by various assays as described in the Examples. Such methods include in vitro assays measuring T cell-mediated killing of target cells bearing the target antigen that is specifically bound by the second antigen-binding polypeptide construct. For example, the killing of target cells bearing the relevant target antigens can be measured in cultures comprising human whole blood, PBMCs, or PBMCs from which the B cells have been removed, referred to herein as "PBMC-B) as a source of effector T cells. Such assays may also be carried out using purified T cell cultures. This type of assay detects both T-cell mediated killing of target cells bearing the target antigen and any killing that occurs through internalization of the drug-conjugated construct by the target cells. Thus in some embodiments described herein, the killing potency of a drug-conjugated antigen-binding construct such as an anti-CD3-CD19, CD3-CDH3, CD3-HER2, CD3-HER3 OR CD3-EGFR against a target cell bearing the target antigen is observed to be higher than the reference unconjugated construct. In some embodiments, the drug-conjugated antigen-binding constructs described herein display increased Raji or Ramos tumor B cell killing compared to a reference unconjugated antigen-binding construct having the same CDRs and binding affinity.

[0186] The direct cytoxicity of a drug-conjugated antigen-binding construct may be determined by culturing the construct with a target cell bearing the target antigen to which the second antigen-binding polypeptide construct is directed. In some embodiments described herein, the target antigen is CD19, HER2, HER3, EGFR or CDH3, and the cytoxicity is determined by culturing the construct with target cells bearing the relevant target antigen, for example, as described in Example 22. In this type of assay, it is possible to assess whether cell killing by means of internalization of the drug, in the absence of any T-cell mediated killing.

[0187] The impact of the drug-conjugated antigen-binding construct on T cells can be measured in several ways. The internalization of the construct into T cells can be measured by coupling a dye or other detectable agent to the construct, and culturing it with T cells and monitoring the amount of dye that accumulates in the T cell, for example, as described in Example 21, in which Jurkat T cells were used. This can be compared in the same experiment with the internalization of the construct into target cells bearing the target antigen. In some embodiments described herein, the internalization of the drug-conjugated construct into T cells is lower than into target cells bearing the target antigen. In some embodiments, the internalization into T cells is at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100-fold lower than internalization into target cells bearing target antigen. In some embodiments, the T cell is a Jurkat cell. In some embodiments the target antigen is EGFR and the construct is internalized at least 10-50-fold more into target cells than T cells. In some embodiments the target antigen is EGFR and the construct is internalized at least 10-50-fold more into target cells than Jurkat T cells. In some embodiments the target antigen is CDH3 and the construct is internalized at least 10-50-fold more into target cells than Jurkat T cells. In some embodiments the target antigen is CD19 and the construct is internalized at least 10-50-fold more into target cells than Jurkat T cells.

[0188] The impact of the drug-conjugated antigen-binding constructs on T cells can also be assessed by culturing T cells from human blood (PBMC) with the constructs, with or without target cells bearing the target antigen, and analyzing the resulting T cell subpopulations in the culture using FACS to detect T cell surface markers PD1, CD4, CD8, CD25, CD69 and CD45. In some embodiments, the assay is carried out as in Examples 14, 15 or 16. In some embodiments, the construct does not increase the number of inhibitory PD1+ cells. In some embodiments, an anti-CD3-CD19-drug conjugate causes less activation of inhibitory (PD-1+) T cells than blinatumomab.

[0189] In some embodiments, the drug-conjugated antigen-binding constructs display killing of Raji or Ramos tumor B cell with high potency, and killing of Jurkat tumor T cells with low potency. In some embodiments, the potency of the drug-conjugated antigen-binding constructs is at least 1.5, 2, 4, 6, 8, 10, 15, 20, 25, 30, 35, 40, 45, 50 or more-fold higher on B cells than on T cells. In a specific embodiment, the potency of a CD3-CD19 antigen-binding construct on Ramos B cells is about 0.5 nM, and the potency on T cells is about 23 nM. In some embodiments, the potency on target cells is at least 2-fold higher than on T cells. In another embodiment, the potency on target cells is at least 4-fold higher than on T cells. In another embodiment, the potency on target cells is at least 6-fold higher than on T cells. In another embodiment, the potency on target cells is at least 8-fold higher than on T cells. In another embodiment, the potency on target cells is at least 10-fold higher than on T cells. In another embodiment, the potency on target cells is at least 15-fold higher than on T cells.

[0190] In some embodiments, the drug-conjugated antigen-binding construct have a cleavable linker and do not reduce the number of T cells in an assay compared to a reference construct with a non-cleavable linker or a reference construct that is not conjugated to a drug.

[0191] The impact of the drug-conjugated antigen-binding constructs on T cells can also be evaluated in vivo, as was done in Examples 18 and 19. In some embodiments, the drug-conjugated antigen-binding constructs did not reduce the number of circulating T cells or the number of splenic T cells in a humanized NSG mouse when administered at doses ranging from 0.1 mg/kg to 3.0 mg/kg. In some embodiments the construct tested was an anti-CD3-CD19-drug conjugate. In some embodiments, the drug-conjugated antigen-binding construct does not substantially impact the level of CD45+/CD8+ T cells in the peripheral blood of humanized NSG mice over a 5-day period.

[0192] In some embodiments, anti-CD3-CD19 antigen-binding constructs described herein are capable of synapse formation and bridging between CD19+ Raji B-cells and Jurkat T-cells as assayed by FACS and/or microscopy. In some embodiments, the drug-conjugated antigen-binding constructs described herein display less activation of inhibitory (PD-1+) T cells than blinatumomab.

[0193] In certain embodiments, the assays are those described in the examples below.

[0194] In some embodiments, the functional characteristics of the bi-specific antigen-binding constructs described herein are compared to those of a reference antigen-binding construct. The identity of the reference antigen-binding construct depends on the functional characteristic being measured or the distinction being made. For example, when comparing the functional characteristics of exemplary anti-CD3-CD19 bi-specific antigen-binding constructs, the reference antigen-binding construct may be the anti CD19 antibody HD37 and/or the anti CD3 antibodies OKT3 or teplizumab. In other embodiment, the reference antigen-binding construct is a construct described herein, e.g., v891 (blinatumomab) or bivalent anti-CD19 (v4371). In some embodiments, the reference antigen-binding construct is the same variant without a conjugated drug, for example, comparing v12043 with v12043 conjugated to DM1 with an SMCC linker.

[0195] The degree to which an antibody blocks binding to a reference antibody, for example, OKT3 or HD37 can be assessed using a competition assay in which the test antibody is able to inhibit or block specific binding of the OKT3 or HD37 antibody (reference antibody) to its target antigen (see, e.g., Junghans et al., Cancer Res. 50:1495, 1990; Fendly et al. Cancer Research 50: 1550-1558; U.S. Pat. No. 6,949,245 for examples of assays). A test antibody competes with a reference antibody if an excess of a test antibody (e.g., at least 2.times., 5.times., 10.times., 20.times., or 100.times.) inhibits or blocks binding of the reference antibody by, e.g., at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% as measured in a competitive binding assay. Test antibodies identified by competition assay (blocking antibodies) include those binding to the same epitope as the reference antibody and antibodies binding to an adjacent epitope sufficiently proximal to the epitope bound by the reference antibody for steric hindrance to occur.

[0196] For example, in one embodiment where one is assaying for the ability of a antigen-binding construct described herein to bind an antigen or to compete with another polypeptide for binding to an antigen, or bind to an Fc receptor and/or anti-albumin antibody, various immunoassays known in the art can be used, including but not limited to, competitive and non-competitive assay systems using techniques such as radioimmunoassays, ELISA (enzyme linked immunosorbent assay), "sandwich" immunoassays, immunoradiometric assays, gel diffusion precipitation reactions, immunodiffusion assays, in situ immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), western blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.

[0197] In certain embodiments, where a binding partner (e.g., a receptor or a ligand) is identified for an antigen-binding domain comprised by a antigen-binding construct described herein, binding to that binding partner by an antigen-binding construct described herein is assayed, e.g., by means well-known in the art, such as, for example, reducing and non-reducing gel chromatography, protein affinity chromatography, and affinity blotting. See generally, Phizicky et al., Microbiol. Rev. 59:94-123 (1995). In another embodiment, the ability of physiological correlates of a antigen-binding construct protein to bind to a substrate(s) of antigen-binding polypeptide constructs of the antigen-binding constructs described herein can be routinely assayed using techniques known in the art.

Antigen-Binding Construct Drug Conjugates (ADCs)

[0198] In many embodiments provided herein antigen binding constructs are conjugated to a drug, e.g., a toxin, a chemotherapeutic agent, a small molecule therapeutic, an immune modulator e.g. a cytokine, or a radioisotope. Numerous methods of preparing ADCs (antibody drug conjugates or antigen binding construct drug conjugates) are known in the art and are described in U.S. Pat. No. 8,624,003 (pot method), U.S. Pat. No. 8,163,888 (one-step), and U.S. Pat. No. 5,208,020 (two-step method) for example.

[0199] In some embodiments, the drug is selected from a maytansine, auristatin, calicheamicin, or derivative thereof. In other embodiments, the drug is a maytansine selected from DM1 and DM4. Further examples are described below.

[0200] In certain embodiments, the antigen binding construct is conjugated to a drug via a linker. The linker may be cleavable or non-cleavable. Non-limiting examples of linkers are described below.

[0201] In some embodiments, one molecule of drug is conjugated to an antigen-binding construct, but in others, multiple drug molecules may be conjugated to the same antigen-binding construct. The drug-to-antigen binding construct ratio (DAR) can be, e.g., in the range of 1.0 to 6.0, or 3.0 to 5.0, or 2.0 to 4.0. In some embodiments described herein, the DAR ranges from 2.2 to 3.5. In some embodiments, the DAR is 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0 or 8.0.

[0202] In certain embodiments, the ADCs have the general formula I:

A-(L-(D).sub.m).sub.n (I)

where A is an antigen binding construct as described herein; L is a linker; D is a drug; m is an integer between 1 and about 10, and n is an integer between 1 and about 20. In certain embodiments, m is between about 1 and about 5, or between 1 and 2. In some embodiments, m is 1. In some embodiments, n is between 1 and 10, for example, between 1 and 8, between 2 and 8, between 2 and 6, or between 2 and 4. In some embodiments, L may be absent.

Drugs

[0203] The drug moiety of the ADCs may be a compound or moiety having a cytostatic or cytotoxic effect. In some embodiments the antigen-binding construct is conjugated to a cytotoxic agent.

[0204] The term "cytotoxic agent" as used herein refers to a substance that inhibits or prevents the function of cells and/or causes destruction of cells. The term is intended to include radioactive isotopes (e.g. .sup.211At, .sup.131I, .sup.125I, .sup.90Y, .sup.186Re, .sup.188Re, .sup.153Sm, .sup.212Bi, .sup.32P and .sup.177Ln), chemotherapeutic agents, and toxins such as small molecule toxins or enzymatically active toxins of bacterial, fungal, plant or animal origin, including fragments and/or variants thereof. One skilled in the art will appreciate that some of these categories of drugs overlap and are thus not intended to be mutually exclusive. For example, toxins may also be considered as chemotherapeutic agents in the sense that they are chemical compounds that may be used to treat cancer. In some embodiments, the drug is an analogue or derivative of a naturally occurring toxin. Examples of such naturally occurring toxins include, but are not limited to, maytansines, auristatins, dolastatins, tubulysins, hemiasterlins, calicheamicins, duocarmycins, pyrrolobenzodiazapenes, amatoxins, camptothecins, Pseudomonas exotoxin (PE), diphtheria toxin (DT), deglycosylated ricin A (dgA) and gelonin. In some embodiments, the drug is an analogue or derivative of a naturally occurring toxin having a peptidyl scaffold. Non-limiting examples of such toxins include auristatins, dolastatins, tubulysins, hemiasterlins and amatoxins.

[0205] In certain embodiments, the drug comprised by the ADCs is a toxin, or a toxin derivative or analogue, where the toxin, derivative or analogue is a microtubule disrupting agent or a DNA modifying agent. Examples of toxins that are microtubule disrupting agents include, but are not limited to, maytansines, auristatins, dolastatins, tubulysins, hemiasterlins, and analogues and derivatives thereof. Examples of toxins that are DNA modifying agents include, but are not limited to, calicheamicins and other enediyne antibiotics, duocarmycins, pyrrolobenzodiazapenes, amatoxins, camptothecins, and analogues and derivatives thereof.

Maytansines

[0206] As indicated above, in some embodiments the drug is a maytansine or maytansine analogue or derivative ("maytansinoid"). Exemplary maytansinoids include DM1 (mertansine, emtansine, N.sub.2'-deacetyl-N.sub.2'-(3-mercapto-1-oxopropyl)maytansine), DM3 (N.sub.2'-deacetyl-N.sub.2'-(4-mercapto-1-oxopentyl)maytansine), and DM4 (ravtansine, soravtansine, N2'-deacetyl-N.sub.2'-(4-methyl-4-mercapto-1-oxopentyl)maytansine) (see U.S. Patent Publication No. US 2009/0202536). Other examples of naturally occurring, synthetic and semi-synthetic maytansinoids are described in Cassady et al., (2004) Chem. Pharm. Bull. 52(1):1-26, and in U.S. Pat. Nos. 4,256,746; 4,361,650; 4,307,016; 4,294,757; 4,424,219; 4,331,598; 4,364,866; 4,313,946; 4,315,929; 4,362,663; 4,322,348 and 4,371,533. Many positions on maytansine compounds are known to be useful as the linkage position, depending upon the type of link. For example, for forming an ester linkage, the C-3 position having a hydroxyl group, the C-14 position modified with hydroxymethyl, the C-15 position modified with a hydroxyl group and the C-20 position having a hydroxyl group are all suitable.

[0207] In certain embodiments, the drug included in the ADC is a maytansinoid having the general formula (II):

##STR00001##

where Y is --(CR.sub.2).sub.m--, each R is independently H or C1-C6 alkyl, m is 1, 2 or 3, and indicates the point of attachment to linker L (see U.S. Pat. No. 5,208,020, RE39151, WO 2007/056550 and Widdison et al., (2006) J. Med. Chem., 49:4392-4408).

[0208] In some embodiments, the drug included in the ADC is a maytansinoid having the general formula (II) in which Y is --CH.sub.2CH.sub.2--, --CH.sub.2CH.sub.2CH(CH.sub.3)-- or --CH.sub.2CH.sub.2C(CH.sub.3).sub.2. All stereoisomers of the maytansine drug moiety are contemplated for the ADCs described herein, i.e. any combination of R and S configurations at the chiral carbons.

[0209] In some embodiments, the drug included in the ADC is a maytansinoid having the following stereochemistry (general formula (IIA)):

##STR00002##

where Y is as defined above for general formula (II).

[0210] In some embodiments, the drug included in the ADC is a maytansinoid having the general formula (II) or (IIA) in which Y is --CH.sub.2CH.sub.2-- (e.g. DM1), --CH.sub.2CH.sub.2CH(CH.sub.3)-- (e.g. DM3) or --CH.sub.2CH.sub.2C(CH.sub.3).sub.2-- (e.g. DM4). In some embodiments, the drug included in the ADC is a maytansinoid having the general formula (II) or (IIA) in which Y is --CH.sub.2CH.sub.2-- (e.g. DM1) or --CH.sub.2CH.sub.2C(CH.sub.3).sub.2-- (e.g. DM4).

Dolastatins and Auristatins

[0211] In some embodiments, the drug is a dolastatin or an auristatin, such as auristatin E (also known in the art as a derivative of dolastatin-10) or auristatin F, or an analogue or derivative thereof. The auristatin can be, for example, an ester formed between auristatin E and a keto acid. For example, auristatin E can be reacted with paraacetyl benzoic acid or benzoylvaleric acid to produce auristatin EB (AEB) and auristatin EVB (AEVB), respectively. Other typical auristatins include auristatin F phenylenediamine (AFP), monomethylauristatin F (MMAF), and monomethylauristatin E (MMAE). The synthesis and structure of exemplary auristatins are described in U.S. Pat. Nos. 6,884,869; 7,098,308; 7,256,257; 7,423,116; 7,498,298 and 7,745,394, each of which is incorporated by reference herein in its entirety and for all purposes.

[0212] The dolastatin or auristatin may be conjugated to the antigen binding construct via the amino (N)-terminus or the carboxy (C)-terminus of the drug molecule. In some embodiments, the drug is an auristatin or analogue or derivative thereof and is conjugated to the antigen binding construct via the N-terminus of the drug molecule. Examples of auristatin analogues suitable for conjugation via the N-terminus of the drug molecule include those described in U.S. Pat. Nos. 7,498,298 and 7,659,241.

[0213] In some embodiments, the drug is MMAE or MMAF. In some embodiments, the drug is MMAE or MMAF and is conjugated to the antigen binding construct via the N-terminus of the drug molecule as shown below, where indicates the point of attachment to linker L:

##STR00003##

[0214] In some embodiments, the drug is an auristatin or analogue or derivative thereof and is conjugated to the antigen binding construct via the C-terminus of the drug molecule. Examples of auristatin analogues suitable for conjugation via the C-terminus of the drug molecule include those described in International Patent Publication Nos. WO 2002/088172 and WO 2016/041082.

[0215] In some embodiments, the drug is an auristatin of general formula (III):

##STR00004##

wherein: R.sup.2 is selected from C.sub.2-C.sub.6 alkyl, aryl, aryl-C.sub.1-C.sub.6 alkyl, C.sub.4-C.sub.7 cycloalkyl, C.sub.3-C.sub.7 cycloalkyl-C.sub.1-C.sub.6 alkyl, heteroaryl, heteroaryl-C.sub.1-C.sub.6 alkyl and heterocyclyl, each optionally substituted with one or more substituents selected from C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkoxycarbonyl, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkylamino, amino, amino-C.sub.1-C.sub.6 alkyl, amino-aryl, amino-C.sub.3-C.sub.7 cycloalkyl, aryl, carboxamide, carboxyl, cyano, C.sub.1-C.sub.6 haloalkyl, C.sub.1-C.sub.6 haloalkoxy, halo, hydroxyl, nitro, thio and thio-C.sub.1-C.sub.6 alkyl; X is --C(O)NHCH(CH.sub.2R.sup.3)--, or X is absent; R.sup.3 is selected from aryl, heteroaryl and C.sub.3-C.sub.7 cycloalkyl, each optionally substituted with one substituent selected from amino and hydroxyl, and R.sup.4 and R.sup.5 are each independently H or C.sub.1-C.sub.6 alkyl.

[0216] In the context of general formula (III), the term "aryl" refers to a radical derived from a 6- to 12-membered mono- or bicyclic hydrocarbon ring system in which at least one ring is aromatic; the term "aryl-alkyl" refers to an alkyl group substituted with one aryl substituent; the term "cycloalkyl-alkyl" refers to an alkyl group substituted with one cycloalkyl substituent; the term "heteroaryl" refers to a radical derived from a 6- to 12-membered mono- or bicyclic ring system wherein at least one ring atom is a heteroatom, such as O, N or S, and at least one ring is aromatic; the term "heteroaryl-alkyl" refers to an alkyl group substituted with one heteroaryl substituent; the term "heterocyclyl" refers to a radical derived from a 3- to 12-membered mono- or bicyclic non-aromatic ring system wherein at least one ring atom is a heteroatom such as O, N or S; the term "alkoxycarbonyl" refers to --C(O)O-alkyl; the term "alkylamino" refers to --NH-alkyl; the term "amino-alkyl" refers to an alkyl group substituted with one amino substituent; the term "amino-aryl" refers to an aryl group substituted with one amino substituent; the term "amino-cycloalkyl" refers to a cycloalkyl group substituted with one amino substituent; the term "carboxamide" refers to --C(O)NH.sub.2; the term "haloalkyl" refers to an alkyl group substituted with one or more halo substituents; the term "haloalkoxy" refers to --O-haloalkyl, and the term "thio-alkyl" refers to --S-alkyl.

[0217] In certain embodiments, the drug is an auristatin of general formula (III) and is conjugated to the antigen binding moiety via the R.sup.2 group.

Tubulysins

[0218] In some embodiments, the drug is a tubulysin. Naturally occurring tubulysins include, for example, tubulysins A, B, C, D, E, F, G, H, I, U, V, W and Z:

##STR00005##

Tubulysin A: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH(CH.sub.3).sub.2; R.sup.3.dbd.OH Tubulysin B: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH.sub.2CH.sub.2; R.sup.3.dbd.OH Tubulysin C: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH.sub.3; R.sup.3.dbd.OH Tubulysin D: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH(CH.sub.3).sub.2; R.sup.3.dbd.H Tubulysin E: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH.sub.2CH.sub.2; R.sup.3.dbd.H Tubulysin F: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH.sub.3; R.sup.3.dbd.H Tubulysin G: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.dbd.C(CH.sub.3).sub.2; R.sup.3.dbd.OH Tubulysin H: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.3; R.sup.3.dbd.H Tubulysin I: R.sup.1.dbd.Ac; R.sup.2.dbd.CH.sub.2OC(O) CH.sub.3; R.sup.3.dbd.OH Tubulysin U: R.sup.1.dbd.Ac; R.sup.2.dbd.R.sup.3.dbd.H Tubulysin V: R.sup.1.dbd.R.sup.2.dbd.R.sup.3.dbd.H Tubulysin W: R.sup.1.dbd.H; R.sup.2.dbd.CH.sub.2OC(O)CH.sub.2CH.sub.2CH.sub.2; R.sup.3.dbd.OH Tubulysin X: R.sup.1.dbd.Ac; R.sup.2.dbd.H; R.sup.3.dbd.OH Tubulysin Z: R.sup.1.dbd.R.sup.2.dbd.H; R.sup.3.dbd.OH

[0219] Therapeutically useful analogues and derivatives of tubulysins have also been described (see, for example, International Patent Publication No. WO 2014/126836 and U.S. Patent Publication No. US 2016/0130299).

[0220] The tubulysin or tubulysin analogue or derivative may be conjugated to the antigen binding construct through a free hydroxyl group, or it may be modified to include an amine group that can be used for conjugation as described in U.S. Patent Publication US 2016/0130299.

Hemiasterlins

[0221] In some embodiments, the drug is a hemiasterlin or analogue or derivative thereof. Various analogues and derivatives of hemiasterlin having anti-mitotic activity have been described (see, for example, International Patent Publication Nos. WO 1996/33211 and WO 2004/026293). U.S. Pat. No. 7,579,323 describes an analogue of hemiasterlin, referred to as HTI-286, that possesses potent anti-mitotic activity and which has been assessed in clinical trials for the treatment of cancer. In certain embodiments, the drug is HTI-286 or a derivative thereof:

##STR00006##

[0222] Additional examples of hemiasterlin analogues are described in International Patent Publication No. WO 2014/144871.

[0223] In certain embodiments, the drug is a hemiasterlin analogue or derivative having general formula (IV):

##STR00007##

wherein: R.sub.26 is selected from optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl; R.sub.27 is selected from optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl; R.sub.16 and R.sub.17 are each independently H or C.sub.1-6 alkyl, and R.sub.18 is C.sub.1-6 alkyl or --SH.

[0224] In the context of general formula (IV), the term "alkyl" refers to a straight or branched chain substituent consisting solely of carbon and hydrogen atoms, which is saturated or unsaturated and has from one to 12 carbon atoms; the term "alkylamino" refers to a substituent of the formula --NHR.sub.a or --NR.sub.aR.sub.a, where each R.sub.a is independently an alkyl substituent containing one to 12 carbon atoms; the term "cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon substituent consisting solely of carbon and hydrogen atoms, which may include fused or bridged ring systems, having from 3 to 10 carbon atoms; the term "aryl" refers to a hydrocarbon ring substituent comprising hydrogen, 6 to 18 carbon atoms and at least one aromatic ring; the term "heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring substituent which consists of 2 to 12 carbon atoms and from one to 6 heteroatoms selected from N, O and S, and the term "heteroaryl" refers to a 5- to 14-membered ring system substituent comprising hydrogen atoms, one to 13 carbon atoms, one to 6 heteroatoms selected from N, O and S, and at least one aromatic ring.

[0225] In certain embodiments, the drug is a hemiasterlin of general formula (IV) and is conjugated to the antigen binding construct via the R.sub.26 substituent. In some embodiments, the drug is a hemiasterlin of general formula (IV) and is conjugated to the antigen binding construct via the R.sub.27 substituent.

Calicheamicins

[0226] In some embodiments, the drug is a calicheamycin or analogue or derivative thereof. Various analogues and derivatives of calicheamycin suitable for conjugation to an antigen binding construct have been described (see, for example, International Patent Publication No. WO 2015/063680, U.S. Pat. Nos. 5,773,001; 5,714,586 and 5,770,701).

Duocarmycins

[0227] In some embodiments, the drug is a duocarmycin or analogue or derivative thereof. Naturally-occurring duocarmycins include, for example, duocarmycins A, B1, B2, C1, C2, D and SA, as well as CC-1065. Various analogues and derivatives of duocarmycins have been described, including adozelesin, bizelesin and centanamycin. Other analogues and derivatives are described in U.S. Pat. Nos. 4,912,227; 5,070,092; 5,084,468; 5,332,837; 5,641,780; 5,739,350 and 8,889,868. Various groups on the duocarmycin molecule may be modified to allow for conjugation to an antigen binding construct. A non-limiting example is provided in Elgersma et al., (2015) Mol. Pharmaceutics, 12:1813-1835.

Pyrrolobenzodiazapenes

[0228] In some embodiments, the drug is a pyrrolobenzodiazapene (PBD) or an analogue or derivative thereof, such as a PBD dimer. Various PBD dimers have been described including, for example, those described in U.S. Pat. Nos. 6,884,799; 7,049,311; 7,511,032; 7,528,126; 7,557,099 and 9,056,914. In some embodiments, the drug is a PBD dimer or an analogue or derivative thereof. The PBD dimer structure is believed to improve the fit at the binding site of DNA. PBD dimers may be conjugated to the antigen binding construct through one of a number of potential linkage sites on the PBD dimer, such as the five-membered pyrrolo ring, the tether between the PBD units, the N10-C11 imine group or the C2 position (see, for example, International Patent Publication Nos. WO 2007/085930, WO 2009/016516, WO 2011/130598, WO 2011/130613 and WO 2011/130616; U.S. Patent Publication No. US 2011/0256157).

Amatoxins

[0229] In some embodiments, the drug is an amatoxin, such as .alpha.-Amanitin, .beta.-Amanitin, .gamma.-Amanitin or .epsilon.-Amanitin, or an analogue or derivative thereof. In some embodiments, the drug is .alpha.-Amanitin or an analogue or derivative thereof. Amatoxins are cyclic peptides composed of eight amino acids and thus present a number of potential sites for conjugation. Various amatoxins and analogues thereof have been described (see, for example, European Patent No. EP 1 859 811, U.S. Pat. No. 9,233,173 and International Patent Publication No. WO 2014/043403).

Camptothecins

[0230] In some embodiments, the drug is a camptothecin (CPT) or analogue or derivative thereof, such as irinotecan (CPT-11), SN-38 (7-ethyl-10-hydroxy-camptothecin), 10-hydroxy camptothecin, topotecan, lurtotecan, 9-aminocamptothecin or 9-nitrocamptothecin. Other examples of CPT analogues and derivatives include 7-butyl-10-amino-camptothecin and 7-butyl-9-amino-10,11-methylenedioxy-camptothecin (see U.S. Patent Publication No. US 2005/0209263) and aniline containing derivatives of these compounds as described in Burke et al., (2009), Bioconj. Chem. 20(6):1242-1250. Conjugation of camptothecin and its analogues or derivatives to the antigen binding construct may be achieved via modification of various groups in the drug molecule. Non-limiting examples are provided in Burke et al., (2009), Bioconj. Chem. 20(6):1242-1250 and Sharkey et al., (2012) Mol. Cancer Ther. 11:224-234.

Chemotherapeutic Agents

[0231] In some embodiments the antigen binding construct is conjugated to a chemotherapeutic agent. Examples include but are not limited to cisplatin and Lapatinib. A "chemotherapeutic agent" is a chemical compound useful in the treatment of cancer.

[0232] Examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN.TM.); alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphaoramide and trimethylolomelamine; nitrogen mustards such as chlorambucil, chlornaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfornithine; elliptinium acetate; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK7; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2',2',2'-trichlorotriethylamine; urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; taxanes, e.g. paclitaxel (TAXOL.RTM., Bristol-Myers Squibb Oncology, Princeton, N.J.) and doxetaxel (TAXOTERE.RTM., Rhone-Poulenc Rorer, Antony, France); chlorambucil; gemcitabine; 6-thioguanine; platinum analogs such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT-11; topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO); retinoic acid; esperamicins; capecitabine; and pharmaceutically acceptable salts, acids or derivatives of any of the above. Also included in this definition are anti-hormonal agents that act to regulate or inhibit hormone action on tumors such as anti-estrogens including for example tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (Fareston); and anti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above.

[0233] In some embodiments, the chemotherapeutic agent is an anthracycline, such as doxorubicin, epirubicin, idarubicin, daunorubicin (also known as daunomycin), nemorubicin or an analogue or derivative thereof. Various groups within the anthracycline molecule may be modified for conjugation to the antigen binding construct. For example, derivatization of daunorubicin and doxorubicin for conjugation to antibodies has been described (see, for example, Kratz et al., (2006) Current Med. Chem. 13:477-523; U.S. Pat. No. 6,630,579).

Linkers

[0234] In some embodiments, the drug is linked to the antigen binding construct, e.g., antibody, by a linker. Linkers are bifunctional or multifunctional moieties capable of linking one or more drugs to the antigen binding construct. In some embodiments, the linker may be bifunctional (or monovalent) such that it links a single drug to a single site on the antigen binding construct. In some embodiments, the linker may be multifunctional (or polyvalent) such that it links more than one drug to a single site on the antigen binding construct. Multifunctional linkers may also be used to link one drug to more than one site on the antigen binding construct in some embodiments.

[0235] Attachment of a linker to an antibody or other antigen binding construct can be accomplished in a variety of ways, such as through surface lysines, reductive-coupling to oxidized carbohydrates, and through cysteine residues liberated by reducing interchain disulfide linkages. Alternatively, attachment of a linker to an antigen binding construct may be achieved by modification of the antigen binding construct to include additional cysteine residues (see, for example, U.S. Pat. Nos. 7,521,541; 8,455,622 and 9,000,130) or non-natural amino acids that provide reactive handles, such as selenomethionine, p-acetylphenylalanine, formylglycine or p-azidomethyl-L-phenylalanine (see, for example, Hofer et al., (2009) Biochemistry 48:12047-12057; Axup et al., (2012) PNAS 109:16101-16106; Wu et al., (2009) PNAS 106:3000-3005; Zimmerman et al., (2014) Bioconj. Chem. 25:351-361), to allow for site-specific conjugation.

[0236] The linkers include a functional group capable of reacting with the target group or groups on the antigen binding construct and one or more functional groups capable of reacting with a target group on the drug. Suitable functional groups are known in the art and include those described, for example, in Bioconjugate Techniques (G. T. Hermanson, 2013, Academic Press). Non-limiting examples of functional groups for reacting with free cysteines or thiols include maleimide, haloacetamide, haloacetyl, activated esters such as succinimide esters, 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters, anhydrides, acid chlorides, sulfonyl chlorides, isocyanates, and isothiocyanates. Also useful in this context are "self-stabilizing" maleimides as described in Lyon et al., (2014) Nat. Biotechnol. 32:1059-1062. Non-limiting examples of functional groups for reacting with surface lysines and amines include activated esters such as N-hydroxysuccinamide (NHS) esters or sulfo-NHS esters, imido esters such as Traut's reagent, isothiocyanates, aldehydes and acid anhydrides such as diethylenetriaminepentaacetic anhydride (DTPA). Other examples include succinimido-1,1,3,3-tetra-methyluronium tetrafluoroborate (TSTU) and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP). Non-limiting examples of functional groups capable of reacting with an electrophilic group on the antigen binding construct or drug (such as an aldehyde or ketone carbonyl group) include hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate and arylhydrazide.

[0237] In certain embodiments, a linker that includes a functional group that allows for bridging of two interchain cysteines on the antibody binding construct may be used, such as a ThioBridge.TM. linker (Badescu et al., (2014) Bioconjug. Chem. 25:1124-1136), a dithiomaleimide (DTM) linker (Behrens et al., 2015, Mol. Pharm. 12:3986-3998), a dithioaryl(TCEP)pyridazinedione based linker (Lee et al., (2016) Chem. Sci. 7:799-802) or a dibromopyridazinedione based linker (Maruani et al., (2015) Nat. Commun. 6:6645).

[0238] A variety of linkers for linking drugs to antibodies and other antigen binding constructs are known in the art, including hydrazone-, disulfide- and peptide-based linkers.

[0239] Suitable linkers typically are more chemically stable to conditions outside the cell than to conditions inside the cell, although less stable linkers may be contemplated in certain situations, such as when the drug is selective or targeted and has a low toxicity to normal cells. Suitable linkers include, for example, cleavable and non-cleavable linkers. A cleavable linker is typically susceptible to cleavage under intracellular conditions, for example, through lysosomal processes. Examples include linkers that are protease-sensitive, acid-sensitive or reduction-sensitive. Non-cleavable linkers by contrast, rely on the degradation of the antibody in the cell, which typically results in the release of an amino acid-linker-cytotoxin moiety.

[0240] Suitable cleavable linkers include, for example, peptide-containing linkers cleavable by an intracellular protease, such as lysosomal protease or an endosomal protease. In exemplary embodiments, the linker can be a dipeptide-containing linker, such as a valine-citrulline (Val-Cit) or a phenylalanine-lysine (Phe-Lys) linker. Other examples of suitable dipeptides for inclusion in the linkers include Val-Lys, Ala-Lys, Phe-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Arg, Ala-Phe, Val-Ala, Met-Lys, Asn-Lys, Ile-Pro, Ile-Val, Asp-Val, His-Val, Met-(D)Lys, Asn-(D)Lys, Val-(D)Asp, NorVal-(D)Asp, Ala-(D)Asp, Me.sub.3Lys-Pro, PhenylGly-(D)Lys, Met-(D)Lys, Asn-(D)Lys, Pro-(D)Lys and Met-(D)Lys. Linkers may also include longer peptide sequences in some embodiments, such as the tripeptides Met-Cit-Val, Gly-Cit-Val, (D)Phe-Phe-Lys or (D)Ala-Phe-Lys, or the tetrapeptides Gly-Phe-Leu-Gly or Ala-Leu-Ala-Leu.

[0241] Additional suitable cleavable linkers include disulfide-containing linkers. Examples of disulfide-containing linkers include, but are not limited to, N-succinimydyl-4-(2-pyridyldithio) butanoate (SPBD) and N-succinimydyl-4-(2-pyridyldithio)-2-sulfo butanoate (sulfo-SPBD). Disulfide-containing linkers may optionally include additional groups to provide steric hindrance adjacent to the disulfide bond in order to improve the extracellular stability of the linker, for example, inclusion of a geminal dimethyl group. Other suitable linkers include linkers hydrolyzable at a specific pH or within a pH range, such as hydrazone linkers. Linkers comprising combinations of these functionalities may also be useful, for example, linkers comprising both a hydrazone and a disulfide are known in the art.

[0242] A further example of a cleavable linker is a linker comprising a .beta.-glucuronide, which is cleavable by .beta.-glucuronidase, an enzyme present in lysosomes and tumor interstitium (see, for example, De Graaf et al., (2002) Curr. Pharm. Des. 8:1391-1403).

[0243] Cleavable linkers may optionally further comprise one or more additional functionalities such as self-immolative and self-elimination groups, stretchers or hydrophilic moieties.

[0244] Self-immolative and self-elimination groups that find use in linkers include, for example, p-aminobenzyloxycarbonyl (PABC) and p-aminobenzyl ether (PABE) groups, and methylated ethylene diamine (MED). Other examples of self-immolative groups include, but are not limited to, aromatic compounds that are electronically similar to the PABC or PABE group such as heterocyclic derivatives, for example 2-aminoimidazol-5-methanol derivatives as described in U.S. Pat. No. 7,375,078. Other examples include groups that undergo cyclization upon amide bond hydrolysis, such as substituted and unsubstituted 4-aminobutyric acid amides (Rodrigues et al. (1995) Chemistry Biology 2:223-227) and 2-aminophenylpropionic acid amides (Amsberry, et al. (1990) J Org. Chem. 55:5867-5877). Self-immolative/self-elimination groups, alone or in combination are often included in peptide-based linkers, but may also be included in other types of linkers. In some embodiments, the linker may include one or more self-immolative and self-elimination groups, for example, a PABC group, a PABE group, or a combination of a PABC or PABE group and an MED.

[0245] Stretchers that find use in linkers for ADCs include, for example, alkylene groups and stretchers based on aliphatic acids, diacids, amines or diamines, such as diglycolate, malonate, caproate and caproamide. Other stretchers include, for example, glycine based stretchers and polyethylene glycol (PEG) or monomethoxy polyethylene glycol (mPEG) stretchers. PEG and mPEG stretchers also function as hydrophilic moieties and may be particularly useful with hydrophobic drugs, although their use in linkers with other drugs is also contemplated in some embodiments.

[0246] In certain embodiments, the linker included in the ADCs of the present disclosure are peptide-based linkers of general formula (VI):

##STR00008##

wherein: Z is a functional group capable of reacting with the target group on the antigen binding construct; Str is a stretcher; AA.sub.1 and AA.sub.2 are each independently an amino acid, wherein AA.sub.1-[AA.sub.2].sub.m forms a protease cleavage site; X is a self-immolative group; D is a drug; n is 0 or 1; m is 1, 2 or 3, and o is 0, 1 or 2.

[0247] In some embodiments, in general formula (VI):

Z is

##STR00009##

[0249] In some embodiments, in general formula (VI):

Str is

##STR00010##

[0250] wherein R is H or C.sub.1-C.sub.6 alkyl; p is an integer between 2 and 10, and q is an integer between 1 and 10.

[0251] In some embodiments, in general formula (VI):

Str is

##STR00011##

[0252] wherein p and q are as defined above. In some embodiments, in general formula (VI):

Str is

##STR00012##

[0253] wherein p is an integer between 2 and 6, and q is an integer between 2 and 8.

[0254] In some embodiments, in general formula (VI):

AA.sub.1-[AA.sub.2].sub.m is selected from Val-Lys, Ala-Lys, Phe-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Arg, Ala-Phe, Val-Ala, Met-Lys, Asn-Lys, Ile-Pro, Ile-Val, Asp-Val, His-Val, Met-(D)Lys, Asn-(D)Lys, Val-(D)Asp, NorVal-(D)Asp, Ala-(D)Asp, Me.sub.3Lys-Pro, PhenylGly-(D)Lys, Met-(D)Lys, Asn-(D)Lys, Pro-(D)Lys, Met-(D)Lys, Met-Cit-Val, Gly-Cit-Val, (D)Phe-Phe-Lys, (D)Ala-Phe-Lys, Gly-Phe-Leu-Gly and Ala-Leu-Ala-Leu.

[0255] In some embodiments, in general formula (VI):

m is 1 (i.e. AA.sub.1-[AA.sub.2].sub.m is a dipeptide).

[0256] In some embodiments, in general formula (VI):

AA.sub.1-[AA.sub.2].sub.m is a dipeptide selected from Val-Lys, Ala-Lys, Phe-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit and Trp-Cit.

[0257] In some embodiments, in general formula (VI):

each X is independently selected from p-aminobenzyloxycarbonyl (PABC), p-aminobenzyl ether (PABE) and methylated ethylene diamine (MED).

[0258] In some embodiments, in general formula (VI):

n is 1.

[0259] In some embodiments, in general formula (VI):

o is 1 or 2.

[0260] In some embodiments, in general formula (VI):

Z is

##STR00013##

[0261] Str is

##STR00014##

[0262] wherein p is an integer between 2 and 6, and q is an integer between 2 and 8; m is 1 and AA.sub.1-[AA.sub.2].sub.m is a dipeptide selected from Val-Lys, Ala-Lys, Phe-Lys, Val-Cit, Phe-Cit, Leu-Cit, Ile-Cit and Trp-Cit; each X is independently selected from p-aminobenzyloxycarbonyl (PABC), p-aminobenzyl ether (PABE) and methylated ethylene diamine (MED). n is 1, and o is 1 or 2.

[0263] In some embodiments, the linker is a disulfide-containing linker and the ADC has general formula (VII):

##STR00015##

wherein: A is the antigen binding construct; D is the drug; Y is --(CH.sub.2).sub.p-- or --(CH.sub.2CH.sub.2O).sub.q--, wherein p and q are each independently an integer between 1 and 10; each R is independently H or C.sub.1-C.sub.6 alkyl; n is 1, 2 or 3, and wherein

##STR00016##

represents an amide bond formed between the linker and the .epsilon.-amino group of a surface lysine on the antigen binding construct.

[0264] In some embodiments in general formula (VII):

p and q are each independently an integer between 1 and 4.

[0265] In some embodiments in general formula (VII):

Y is --(CH.sub.2).sub.p-- and p is an integer between 1 and 4.

[0266] In some embodiments in general formula (VII):

each R is independently H or Me.

[0267] In some embodiments in general formula (VII):

n is 1 or 2.

[0268] In some embodiments in general formula (VII):

Y is --(CH.sub.2).sub.p-- and p is an integer between 1 and 4; each R is independently H or Me, and n is 1 or 2.

[0269] Examples of commonly used cleavable linkers that may find use in the ADCs of the present disclosure in some embodiments include, but are not limited to, linkers comprising SPBD, sulfo-SPBD, hydrazone, Val-Cit, maleidocaproyl (MC or mc), mc-Val-Cit, mc-Val-Cit-PABC, Phe-Lys, mc-Phe-Lys or mc-Phe-Lys-PABC.

[0270] Various non-cleavable linkers are known in the art for linking drugs to targeting moieties and may be useful in the ADCs of the present disclosure. Examples of non-cleavable linkers include linkers having an N-succinimidyl ester or N-sulfosuccinimidyl ester moiety for reaction with the cell binding agent, as well as a maleimido- or haloacetyl-based moiety for reaction with the drug, or vice versa. An example of such a non-cleavable linker is based on sulfosuccinimidyl-4-[N-maleimidomethyl]cyclohexane-1-carboxylate (sulfo-SMCC). Sulfo-SMCC conjugation typically occurs via a maleimide group which reacts with sulfhydryls (thiols, --SH) on the drug moiety, while the sulfo-NHS ester is reactive toward primary amines (as found in lysine and the protein or peptide N-terminus). Other non-limiting examples of such linkers include those based on N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidocaproa- te) ("long chain" SMCC or LC-SMCC), K-maleimidoundecanoic acid N-succinimidyl ester (KMUA), .gamma.-maleimidobutyric acid N-succinimidyl ester (GMBS), .epsilon.-maleimidocaproic acid N-hydroxysuccinimide ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(.alpha.-maleimidoacetoxy)-succinimide ester (AMAS), succinimidyl-6-(.beta.-maleimidopropionamido)hexanoate (SMPH), N-succinimidyl 4-(p-maleimidophenyl)-butyrate (SMPB), and N-(p-maleimidophenyl)isocyanate (PMPI). Other examples include those comprising a haloacetyl-based functional group such as N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB), N-succinimidyl iodoacetate (SIA), N-succinimidyl bromoacetate (SBA) and N-succinimidyl 3-(bromoacetamido)propionate (SBAP).

[0271] Other examples of non-cleavable linkers include maleimidocarboxylic acids, such as maleimidocaproyl (MC).

[0272] In certain embodiments, the antigen binding construct is conjugated to the drug via a sulphonamide-containing linker as described in International Patent Publication No. WO 2015/095953. In some embodiments, the antigen binding construct is conjugated to the drug via a linker having general formula (VIII):

##STR00017##

wherein: R is selected from optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heteroaryl, --COR.sup.27--, --CSR.sup.27--, --OR.sup.27-- and --NHR.sup.27--, wherein each R.sup.27 is independently selected from optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl; P.sup.3 is the drug or a portion of the drug; L.sup.3 is a linker or a portion of a linker, and A is the antigen binding construct.

[0273] In some embodiments, the antigen binding construct is conjugated to the drug via a linker having general formula (IX):

##STR00018##

wherein -L.sup.3-A has the structure:

##STR00019##

wherein: P.sup.3 is a remaining portion of the drug; the --NH-- group bonded to R' forms a peptide bond (the junction peptide bond or JPB) with (AA)'; R' is selected from optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heteroaryl, --COR.sup.27--, --CSR.sup.27--, --OR.sup.27-- and --NHR.sup.27--, wherein each R.sup.27 is independently selected from optionally substituted alkyl, optionally substituted alkylamino, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl and optionally substituted heteroaryl; each AA is independently an amino acid, wherein (AA).sup.1-(AA).sub.x taken together comprise an amino acid sequence capable of facilitating cleavage of the JPB; x is an integer from 0 to 25; L' is a remaining portion of the linker or is absent; A is the antigen binding construct.

[0274] In some embodiments, the antigen-binding construct is coupled to the drug via a cleavable linker e.g. a SPBD linker or a maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-Val-Cit-PABC) linker. In some embodiments, the antigen-binding construct is coupled to the drug via a non-cleavable linker e.g. a MCC linker formed using SMCC or sulfo-SMCC.

[0275] Selection of an appropriate linker for a given ADC may be readily made by the skilled person having knowledge of the art and taking into account relevant factors, such as the site of attachment to the antigen binding construct, any structural constraints of the drug and the hydrophobicity of the drug (see, for example, review in Nolting, Chapter 5, Antibody-Drug Conjugates: Methods in Molecular Biology, 2013, Ducry (Ed.), Springer). A number of specific linker-toxin combinations have been described and may be used with the antigen binding constructs described herein to prepare ADCs in certain embodiments. Examples include, but are not limited to, cleavable peptide-based linkers with auristatins such as MMAE and MMAF, camptothecins such as SN-38, duocarmycins and PBD dimers; non-cleavable MC-based linkers with auristatins MMAF and MMAE; acid-labile hydrazone-based linkers with calicheamicins and doxorubicin; disulfide-based linkers with maytansinoids such as DM1 and DM4, and bis-maleimido-trioxyethylene glycol (BMPEO)-based linkers with maytansinoid DM1 (see, for example, Peters & Brown, (2015) Biosci. Rep. e00225; Dosio et al., (2014) Recent Patents on Anti-Cancer Drug Discovery 9:35-65; US Patent Publication No. US 2015/0374847).

Preparation of ADCs

[0276] The ADC may be prepared by one of several routes known in the art, employing organic chemistry reactions, conditions, and reagents known to those skilled in the art (see, for example, Bioconjugate Techniques (G. T. Hermanson, 2013, Academic Press). For example, conjugation may be achieved by (1) reaction of a nucleophilic group or an electrophilic group of an antibody with a bivalent linker reagent, to form antibody-linker intermediate Ab-L, via a covalent bond, followed by reaction with an activated drug moiety D; or (2) reaction of a nucleophilic group or an electrophilic group of a drug moiety with a linker reagent, to form drug-linker intermediate D-L, via a covalent bond, followed by reaction with the nucleophilic group or an electrophilic group of an antibody. Conjugation methods (1) and (2) may be employed with a variety of antibodies, drug moieties, and linkers to prepare the ADCs described here. Various prepared linkers, linker components and toxins are commercially available or may be prepared using standard synthetic organic chemistry techniques (see, for example, March's Advanced Organic Chemistry (Smith & March, 2006, Sixth Ed., Wiley); Toki et al., (2002) J. Org. Chem. 67:1866-1872; Frisch et al., (1997) Bioconj. Chem. 7:180-186; Bioconjugate Techniques (G. T. Hermanson, 2013, Academic Press)). In addition, a number of pre-formed drug-linkers suitable for reaction with a selected antigen binding construct are also available commercially, for example, linker-toxins comprising DM1, DM4, MMAE, MMAF or Duocarmycin SA are available from Creative BioLabs (Shirley, N.Y.).

[0277] Several specific examples of methods of preparing ADCs are known in the art and are described in U.S. Pat. No. 8,624,003 (pot method), U.S. Pat. No. 8,163,888 (one-step), and U.S. Pat. No. 5,208,020 (two-step method). Other methods are known in the art and include those described in Antibody-Drug Conjugates: Methods in Molecular Biology, 2013, Ducry (Ed.), Springer. In addition, various antibody drug conjugation services are available commercially from companies such as Lonza Inc. (Allendale, N.J.), Abzena PLC (Cambridge, UK), ADC Biotechnology (St. Asaph, UK), Baxter BioPharma Solutions (Baxter Healthcare Corporation, Deerfield, Ill.) and Piramel Pharma Solutions (Grangemouth, UK).

[0278] The average number of drugs conjugated to the antigen binding construct (drug-to-antibody ratio or DAR) may be determined by standard techniques such as UV/VIS spectroscopic analysis, ELISA-based techniques, chromatography techniques such as hydrophobic interaction chromatography (HIC), UV-MALDI mass spectrometry (MS) and MALDI-TOF MS. In addition, distribution of drug-linked forms (for example, the fraction of antigen binding constructs containing zero, one, two, three, etc. drugs) may also optionally be analyzed. Various techniques are known in the art to measure such distribution, including MS (with or without an accompanying chromatographic separation step), hydrophobic interaction chromatography, reverse-phase HPLC or iso-electric focusing gel electrophoresis (IEF) (see, for example, Wakankar et al., (2011) mAbs 3:161-172).

Pharmaceutical Compositions

[0279] Also provided herein are pharmaceutical compositions comprising a drug-conjugated antigen-binding construct described herein. Pharmaceutical compositions comprise the construct and a pharmaceutically acceptable carrier.

[0280] The term "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. In some aspects, the carrier is a man-made carrier not found in nature. Water can be used as a carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. Oral formulation can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin. Such compositions will contain a therapeutically effective amount of the compound, preferably in purified form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

[0281] In certain embodiments, the composition comprising the construct is formulated in accordance with routine procedures as a pharmaceutical composition adapted for intravenous administration to human beings. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. Where necessary, the composition may also include a solubilizing agent and a local anaesthetic such as lignocaine to ease pain at the site of the injection. Generally, the ingredients are supplied either separately or mixed together in unit dosage form, for example, as a dry lyophilized powder or water free concentrate in a hermetically sealed container such as an ampoule or sachette indicating the quantity of active agent. Where the composition is to be administered by infusion, it can be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. Where the composition is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients may be mixed prior to administration.

[0282] In certain embodiments, the compositions described herein are formulated as neutral or salt forms. Pharmaceutically acceptable salts include those formed with anions such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with cations such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxide isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

Methods of Treatment

[0283] Also described herein are methods of treating a disease or disorder comprising administering to a subject in which such treatment, prevention or amelioration is desired, an antigen-binding construct described herein, in an amount effective to treat, prevent or ameliorate the disease or disorder.

[0284] Disorder and disease are used interchangeably and refer to any condition that would benefit from treatment with an antigen-binding construct or method described herein. This includes chronic and acute disorders or diseases including those pathological conditions which predispose the mammal to the disorder in question. In some embodiments, the disorder is cancer.

[0285] The term "subject" refers to an animal which is the object of treatment, observation or experiment. An animal may be a human, a non-human primate, a companion animal (e.g., dogs, cats, and the like), farm animal (e.g., cows, sheep, pigs, horses, and the like) or a laboratory animal (e.g., rats, mice, guinea pigs, and the like).

[0286] The term "mammal" as used herein includes but is not limited to humans, non-human primates, canines, felines, murines, bovines, equines, and porcines.

[0287] "Treatment" refers to clinical intervention in an attempt to alter the natural course of the individual or cell being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include preventing occurrence or recurrence of disease, alleviation of symptoms, diminishing of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. In some embodiments, antigen-binding constructs described herein are used to delay development of a disease or disorder. In one embodiment, antigen-binding constructs and methods described herein effect tumor regression. In one embodiment, antigen-binding constructs and methods described herein effect inhibition of tumor/cancer growth.

[0288] Desirable effects of treatment include, but are not limited to, preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. In some embodiments, construct constructs described herein are used to delay development of a disease or to slow the progression of a disease.

[0289] The term "effective amount" as used herein refers to that amount of construct being administered, which will accomplish the goal of the recited method, e.g., relieve to some extent one or more of the symptoms of the disease, condition or disorder being treated. The amount of the composition described herein which will be effective in the treatment, inhibition and prevention of a disease or disorder associated with aberrant expression and/or activity of a therapeutic protein can be determined by standard clinical techniques. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the seriousness of the disease or disorder, and should be decided according to the judgment of the practitioner and each patient's circumstances. Effective doses are extrapolated from dose-response curves derived from in vitro or animal model test systems.

Therapeutic Uses:

[0290] In an aspect, the antigen-binding constructs and drug-conjugated antigen-binding constructs described herein are used in antibody-based therapies which involve administering the antigen-binding constructs, or nucleic acids encoding antigen-binding constructs to a patient for treating one or more diseases, disorders, or conditions. Such disorders, diseases and conditions may include, but are not limited to, cancer (hematological, solid tumor or metastatic), autoimmune diseases, inflammatory diseases, and diseases caused by pathogen such as viruses, bacteria, parasites or fungi that express antigens on the cell surface of an infected host. Targets useful in these constructs are found in Table LL.

[0291] In some embodiments, the drug-conjugated antigen-binding constructs do not substantially deplete the T cells of a subject to which the construct is administered. As used herein "substantially deplete" T cells means reduce the number of T cells to a number that is less than about 75%, less than about 50%, or less than about 25 percent of the pre-administration number.

[0292] In certain embodiments is provided a method for the prevention, treatment or amelioration of cancer, said method comprising administering to a subject in need of such prevention, treatment or amelioration a pharmaceutical composition comprising an antigen-binding construct described herein.

[0293] In certain embodiments is a method of treating cancer in a mammal in need thereof, comprising administering to the mammal a composition comprising an effective amount of the pharmaceutical composition described herein, optionally in combination with other pharmaceutically active molecules. In certain embodiments, the cancer is a lymphoma or leukemia.

[0294] In some embodiments, the cancer is a lymphoma or leukemia or a B cell malignancy, or a cancer that expresses CD19, or non-Hodgkin's lymphoma (NHL) or mantle cell lymphoma (MCL) or acute lymphoblastic leukemia (ALL) or chronic lymphocytic leukemia (CLL) or rituximab- or CHOP (Cytoxan.TM./Adriamycin.TM. vincristine/prednisone therapy)-resistant B cell cancer, or a blinatumomab-resistant or refractory B cell cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a non-inflammatory tumor that is not easily infiltrated with lymphocytes.

[0295] In a further aspect, the antigen-binding constructs and drug-conjugated antigen-binding constructs described herein are for use in the manufacture or preparation of a medicament. In one embodiment, the medicament is for treatment of cancer. In certain embodiments, the medicament is for the treatment of lymphoma or leukemia. In other embodiments, the medicament is for the treatment of cancer described above. In another embodiment, the medicament is for use in a method of treating cancer comprising administering to patient having cancer, an effective amount of the medicament.

[0296] In certain embodiments, the methods and uses described herein further comprise administering to the patient an effective amount of at least one additional therapeutic agent, e.g., cytotoxic agents, chemotherapeutic agents, cytokines, growth inhibitory agents, kinase inhibitors, anti-angiogenic agents, cardioprotectants, immunostimulatory agents, immunosuppressive agents, protein tyrosine kinase (PTK) inhibitors, other antibodies, Fc fusions, or immunoglobulins, or other therapeutic agents.

[0297] In certain embodiments, the additional therapeutic agent is for preventing and/or treating cancer. Such combination therapy encompasses combined administration (where two or more therapeutic agents are included in the same or separate formulations), and separate administration, in which case, administration of the antigen-binding construct described herein can occur prior to, simultaneously, and/or following, administration of the additional therapeutic agent and/or adjuvant.

[0298] The antigen-binding constructs and drug-conjugated antigen-binding constructs described herein may be administered alone or in combination with other types of treatments (e.g., radiation therapy, chemotherapy, hormonal therapy, immunotherapy and anti-tumor agents).

[0299] Demonstration of Therapeutic or Prophylactic Activity:

[0300] The drug-conjugated antigen-binding constructs or pharmaceutical compositions described herein are tested in vitro, and then in vivo for the desired therapeutic or prophylactic activity, prior to use in humans. For example, in vitro assays to demonstrate the therapeutic or prophylactic utility of a compound or pharmaceutical composition include, the effect of a compound on a cell line or a patient tissue sample. The effect of the compound or composition on the cell line and/or tissue sample can be determined utilizing techniques known to those of skill in the art including, but not limited to, rosette formation assays and cell lysis assays.

[0301] Therapeutic/Prophylactic Administration and Composition:

[0302] Provided are methods of treatment, inhibition and prophylaxis by administration to a subject of an effective amount of an antigen-binding construct or pharmaceutical composition described herein. In an embodiment, the antigen-binding construct is substantially purified (e.g., substantially free from substances that limit its effect or produce undesired side-effects). In certain embodiments, the subject is an animal, including but not limited to animals such as cows, pigs, horses, chickens, cats, dogs, etc., and in certain embodiments, a mammal, and most preferably human.

[0303] Various delivery systems are known and can be used to administer an antigen-binding construct formulation described herein, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the antigen-binding constructs, receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-4432 (1987)), construction of a nucleic acid as part of a retroviral or other vector, etc. Methods of introduction include but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The antigen-binding constructs may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other therapeutic agents. Administration can be systemic or local. Suitable routes of administration include intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example, attached to a reservoir, such as an Ommaya reservoir. Pulmonary administration can also be employed, e.g., by use of an inhaler or nebulizer, and formulation with an aerosolizing agent.

[0304] In a specific embodiment, it is desirable to administer the antigen-binding constructs, or compositions described herein locally to the area in need of treatment; this may be achieved by, for example, and not by way of limitation, local infusion during surgery, topical application, e.g., in conjunction with a wound dressing after surgery, by injection, by means of a catheter, by means of a suppository, or by means of an implant, said implant being of a porous, non-porous, or gelatinous material, including membranes, such as sialastic membranes, or fibers. Preferably, when administering a protein, including an antibody, of the invention, care must be taken to use materials to which the protein does not absorb.

[0305] In another embodiment, the antigen-binding constructs or composition can be delivered in a vesicle, in particular a liposome (see Langer, Science 249:1527-1533 (1990); Treat et al., in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez-Berestein and Fidler (eds.), Liss, New York, pp. 353-365 (1989); Lopez-Berestein, ibid., pp. 317-327; see generally ibid.)

[0306] In yet another embodiment, the antigen-binding constructs or composition can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton, CRC Crit. Ref. Biomed. Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); Saudek et al., N. Engl. J. Med. 321:574 (1989)). In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J., Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983); see also Levy et al., Science 228:190 (1985); During et al., Ann Neurol. 25:351 (1989); Howard et al., J. Neurosurg. 71:105 (1989)). In yet another embodiment, a controlled release system can be placed in proximity of the therapeutic target, e.g., the brain, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138 (1984)).

[0307] Other controlled release systems are discussed in the review by Langer (Science 249:1527-1533 (1990)).

Kits and Articles of Manufacture

[0308] Also described herein are kits comprising one or more antigen-binding constructs described herein. Individual components of the kit would be packaged in separate containers and, associated with such containers, can be a notice in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals or biological products, which notice reflects approval by the agency of manufacture, use or sale. The kit may optionally contain instructions or directions outlining the method of use or administration regimen for the antigen-binding construct, sometimes referred to as a "package insert".

[0309] When one or more components of the kit are provided as solutions, for example an aqueous solution, or a sterile aqueous solution, the container means may itself be an inhalant, syringe, pipette, eye dropper, or other such like apparatus, from which the solution may be administered to a subject or applied to and mixed with the other components of the kit.

[0310] The components of the kit may also be provided in dried or lyophilized form and the kit can additionally contain a suitable solvent for reconstitution of the lyophilized components. Irrespective of the number or type of containers, the kits described herein also may comprise an instrument for assisting with the administration of the composition to a patient. Such an instrument may be an inhalant, nasal spray device, syringe, pipette, forceps, measured spoon, eye dropper or similar medically approved delivery vehicle.

[0311] In another aspect described herein, an article of manufacture containing materials useful for the treatment, prevention and/or diagnosis of the disorders described above is provided. The article of manufacture comprises a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, IV solution bags, etc. The containers may be formed from a variety of materials such as glass or plastic. The container holds a composition which is by itself or combined with another composition effective for treating, preventing and/or diagnosing the condition and may have a sterile access port (for example the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). At least one active agent in the composition is a T cell activating antigen-binding construct described herein. The label or package insert indicates that the composition is used for treating the condition of choice. Moreover, the article of manufacture may comprise (a) a first container with a composition contained therein, wherein the composition comprises an antigen-binding construct described herein; and (b) a second container with a composition contained therein, wherein the composition comprises a further cytotoxic or otherwise therapeutic agent. The article of manufacture in this embodiment described herein may further comprise a package insert indicating that the compositions can be used to treat a particular condition. Alternatively, or additionally, the article of manufacture may further comprise a second (or third) container comprising a pharmaceutically-acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate-buffered saline, Ringer's solution and dextrose solution. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, and syringes.

Polypeptides and Polynucleotides

[0312] The antigen-binding constructs described herein comprise at least one polypeptide. Also described are polynucleotides encoding the polypeptides described herein. The polypeptides and polynucleotides are typically isolated.

[0313] As used herein, "isolated" means an agent (e.g., a polypeptide or polynucleotide) that has been identified and separated and/or recovered from a component of its natural cell culture environment. Contaminant components of its natural environment are materials that would interfere with diagnostic or therapeutic uses for the antigen-binding construct, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. Isolated also refers to an agent that has been synthetically produced, e.g., via human intervention.

[0314] The terms "polypeptide," "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. That is, a description directed to a polypeptide applies equally to a description of a peptide and a description of a protein, and vice versa. The terms apply to naturally occurring amino acid polymers as well as amino acid polymers in which one or more amino acid residues is a non-naturally encoded amino acid. As used herein, the terms encompass amino acid chains of any length, including full length proteins, wherein the amino acid residues are linked by covalent peptide bonds.

[0315] The term "amino acid" refers to naturally occurring and non-naturally occurring amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally encoded amino acids are the 20 common amino acids (alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, praline, serine, threonine, tryptophan, tyrosine, and valine) and pyrrolysine and selenocysteine. Amino acid analogs refers to compounds that have the same basic chemical structure as a naturally occurring amino acid, i.e., an a carbon that is bound to a hydrogen, a carboxyl group, an amino group, and an R group, such as, homoserine, norleucine, methionine sulfoxide, methionine methyl sulfonium. Such analogs have modified R groups (such as, norleucine) or modified peptide backbones, but retain the same basic chemical structure as a naturally occurring amino acid. Reference to an amino acid includes, for example, naturally occurring proteogenic L-amino acids; D-amino acids, chemically modified amino acids such as amino acid variants and derivatives; naturally occurring non-proteogenic amino acids such as .beta.-alanine, ornithine, etc.; and chemically synthesized compounds having properties known in the art to be characteristic of amino acids. Examples of non-naturally occurring amino acids include, but are not limited to, .alpha.-methyl amino acids (e.g. .alpha.-methyl alanine), D-amino acids, histidine-like amino acids (e.g., 2-amino-histidine, .beta.-hydroxy-histidine, homohistidine), amino acids having an extra methylene in the side chain ("homo" amino acids), and amino acids in which a carboxylic acid functional group in the side chain is replaced with a sulfonic acid group (e.g., cysteic acid). The incorporation of non-natural amino acids, including synthetic non-native amino acids, substituted amino acids, or one or more D-amino acids into the proteins of the present invention may be advantageous in a number of different ways. D-amino acid-containing peptides, etc., exhibit increased stability in vitro or in vivo compared to L-amino acid-containing counterparts. Thus, the construction of peptides, etc., incorporating D-amino acids can be particularly useful when greater intracellular stability is desired or required. More specifically, D-peptides, etc., are resistant to endogenous peptidases and proteases, thereby providing improved bioavailability of the molecule, and prolonged lifetimes in vivo when such properties are desirable. Additionally, D-peptides, etc., cannot be processed efficiently for major histocompatibility complex class II-restricted presentation to T helper cells, and are therefore, less likely to induce humoral immune responses in the whole organism.

[0316] Amino acids may be referred to herein by either their commonly known three letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Nucleotides, likewise, may be referred to by their commonly accepted single-letter codes.

[0317] Also described herein are polynucleotides encoding polypeptides of the antigen-binding constructs. The term "polynucleotide" or "nucleotide sequence" is intended to indicate a consecutive stretch of two or more nucleotide molecules. The nucleotide sequence may be of genomic, cDNA, RNA, semisynthetic or synthetic origin, or any combination thereof.

[0318] The term "nucleic acid" refers to deoxyribonucleotides, deoxyribonucleosides, ribonucleosides, or ribonucleotides and polymers thereof in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides which have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless specifically limited otherwise, the term also refers to oligonucleotide analogs including PNA (peptidonucleic acid), analogs of DNA used in antisense technology (phosphorothioates, phosphoroamidates, and the like). Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (including but not limited to, degenerate codon substitutions) and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxyinosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)).

[0319] "Conservatively modified variants" applies to both amino acid and nucleic acid sequences. With respect to particular nucleic acid sequences, "conservatively modified variants" refers to those nucleic acids which encode identical or essentially identical amino acid sequences, or where the nucleic acid does not encode an amino acid sequence, to essentially identical sequences. Because of the degeneracy of the genetic code, a large number of functionally identical nucleic acids encode any given protein. For instance, the codons GCA, GCC, GCG and GCU all encode the amino acid alanine. Thus, at every position where an alanine is specified by a codon, the codon can be altered to any of the corresponding codons described without altering the encoded polypeptide. Such nucleic acid variations are "silent variations," which are one species of conservatively modified variations. Every nucleic acid sequence herein which encodes a polypeptide also describes every possible silent variation of the nucleic acid. One of ordinary skill in the art will recognize that each codon in a nucleic acid (except AUG, which is ordinarily the only codon for methionine, and TGG, which is ordinarily the only codon for tryptophan) can be modified to yield a functionally identical molecule. Accordingly, each silent variation of a nucleic acid which encodes a polypeptide is implicit in each described sequence.

[0320] As to amino acid sequences, one of ordinary skill in the art will recognize that individual substitutions, deletions or additions to a nucleic acid, peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a "conservatively modified variant" where the alteration results in the deletion of an amino acid, addition of an amino acid, or substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are known to those of ordinary skill in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecies homologs, and alleles described herein.

[0321] Conservative substitution tables providing functionally similar amino acids are known to those of ordinary skill in the art. The following eight groups each contain amino acids that are conservative substitutions for one another: 1) Alanine (A), Glycine (G); 2) Aspartic acid (D), Glutamic acid (E); 3) Asparagine (N), Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V); 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W); 7) Serine (S), Threonine (T); and [0139] 8) Cysteine (C), Methionine (M) (see, e.g., Creighton, Proteins: Structures and Molecular Properties (W H Freeman & Co.; 2nd edition (December 1993)

[0322] The terms "identical" or percent "identity," in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same. Sequences are "substantially identical" if they have a percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% identity over a specified region), when compared and aligned for maximum correspondence over a comparison window, or designated region as measured using one of the following sequence comparison algorithms (or other algorithms available to persons of ordinary skill in the art) or by manual alignment and visual inspection. This definition also refers to the complement of a test sequence. The identity can exist over a region that is at least about 50 amino acids or nucleotides in length, or over a region that is 75-100 amino acids or nucleotides in length, or, where not specified, across the entire sequence of a polynucleotide or polypeptide. A polynucleotide encoding a polypeptide of the present invention, including homologs from species other than human, may be obtained by a process comprising the steps of screening a library under stringent hybridization conditions with a labeled probe having a polynucleotide sequence described herein or a fragment thereof, and isolating full-length cDNA and genomic clones containing said polynucleotide sequence. Such hybridization techniques are well known to the skilled artisan.

[0323] For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.

[0324] A "comparison window", as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are known to those of ordinary skill in the art. Optimal alignment of sequences for comparison can be conducted, including but not limited to, by the local homology algorithm of Smith and Waterman (1970) Adv. Appl. Math. 2:482c, by the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443, by the search for similarity method of Pearson and Lipman (1988) Proc. Nat'l. Acad. Sci. USA 85:2444, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Ausubel et al., Current Protocols in Molecular Biology (1995 supplement)).

[0325] One example of an algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1997) Nuc. Acids Res. 25:3389-3402, and Altschul et al. (1990) J. Mol. Biol. 215:403-410, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information available at the World Wide Web at ncbi.nlm.nih.gov. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) or 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1992) Proc. Natl. Acad. Sci. USA 89:10915) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands. The BLAST algorithm is typically performed with the "low complexity" filter turned off.

[0326] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5787). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, or less than about 0.01, or less than about 0.001.

[0327] The phrase "selectively (or specifically) hybridizes to" refers to the binding, duplexing, or hybridizing of a molecule only to a particular nucleotide sequence under stringent hybridization conditions when that sequence is present in a complex mixture (including but not limited to, total cellular or library DNA or RNA).

[0328] The phrase "stringent hybridization conditions" refers to hybridization of sequences of DNA, RNA, or other nucleic acids, or combinations thereof under conditions of low ionic strength and high temperature as is known in the art. Typically, under stringent conditions a probe will hybridize to its target subsequence in a complex mixture of nucleic acid (including but not limited to, total cellular or library DNA or RNA) but does not hybridize to other sequences in the complex mixture. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures. An extensive guide to the hybridization of nucleic acids is found in Tijssen, Laboratory Techniques in Biochemistry and Molecular Biology--Hybridization with Nucleic Probes, "Overview of principles of hybridization and the strategy of nucleic acid assays" (1993).

[0329] As used herein, the terms "engineer, engineered, engineering", are considered to include any manipulation of the peptide backbone or the post-translational modifications of a naturally occurring or recombinant polypeptide or fragment thereof. Engineering includes modifications of the amino acid sequence, of the glycosylation pattern, or of the side chain group of individual amino acids, as well as combinations of these approaches. The engineered proteins are expressed and produced by standard molecular biology techniques.

[0330] By "isolated nucleic acid molecule or polynucleotide" is intended a nucleic acid molecule, DNA or RNA, which has been removed from its native environment. For example, a recombinant polynucleotide encoding a polypeptide contained in a vector is considered isolated. Further examples of an isolated polynucleotide include recombinant polynucleotides maintained in heterologous host cells or purified (partially or substantially) polynucleotides in solution. An isolated polynucleotide includes a polynucleotide molecule contained in cells that ordinarily contain the polynucleotide molecule, but the polynucleotide molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location. Isolated RNA molecules include in vivo or in vitro RNA transcripts, as well as positive and negative strand forms, and double-stranded forms. Isolated polynucleotides or nucleic acids described herein, further include such molecules produced synthetically, e.g., via PCR or chemical synthesis. In addition, a polynucleotide or a nucleic acid, in certain embodiments, include a regulatory element such as a promoter, ribosome binding site, or a transcription terminator.

[0331] The term "polymerase chain reaction" or "PCR" generally refers to a method for amplification of a desired nucleotide sequence in vitro, as described, for example, in U.S. Pat. No. 4,683,195. In general, the PCR method involves repeated cycles of primer extension synthesis, using oligonucleotide primers capable of hybridising preferentially to a template nucleic acid.

[0332] By a nucleic acid or polynucleotide having a nucleotide sequence at least, for example, 95% "identical" to a reference nucleotide sequence of the present invention, it is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These alterations of the reference sequence may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence. As a practical matter, whether any particular polynucleotide sequence is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence of the present invention can be determined conventionally using known computer programs, such as the ones discussed above for polypeptides (e.g. ALIGN-2).

[0333] A derivative, or a variant of a polypeptide is said to share "homology" or be "homologous" with the peptide if the amino acid sequences of the derivative or variant has at least 50% identity with a 100 amino acid sequence from the original peptide. In certain embodiments, the derivative or variant is at least 75% the same as that of either the peptide or a fragment of the peptide having the same number of amino acid residues as the derivative. In certain embodiments, the derivative or variant is at least 85% the same as that of either the peptide or a fragment of the peptide having the same number of amino acid residues as the derivative. In certain embodiments, the amino acid sequence of the derivative is at least 90% the same as the peptide or a fragment of the peptide having the same number of amino acid residues as the derivative. In some embodiments, the amino acid sequence of the derivative is at least 95% the same as the peptide or a fragment of the peptide having the same number of amino acid residues as the derivative. In certain embodiments, the derivative or variant is at least 99% the same as that of either the peptide or a fragment of the peptide having the same number of amino acid residues as the derivative.

[0334] The term "modified," as used herein refers to any changes made to a given polypeptide, such as changes to the length of the polypeptide, the amino acid sequence, chemical structure, co-translational modification, or post-translational modification of a polypeptide. The form "(modified)" term means that the polypeptides being discussed are optionally modified, that is, the polypeptides under discussion can be modified or unmodified.

[0335] In some aspects, an antigen-binding construct comprises an amino acids sequence that is at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical to a relevant amino acid sequence or fragment thereof set forth in the Table(s) or accession number(s) disclosed herein. In some aspects, an isolated antigen-binding construct comprises an amino acids sequence encoded by a polynucleotide that is at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identical to a relevant nucleotide sequence or fragment thereof set forth in Table(s) or accession number(s) disclosed herein.

[0336] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. In the event that there are a plurality of definitions for terms herein, those in this section prevail. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information. Terms understood by those in the art of antibody technology are each given the meaning acquired in the art, unless expressly defined differently herein.

[0337] It is to be understood that the general description and following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed.

[0338] In this application, the use of the singular includes the plural unless specifically stated otherwise.

[0339] In the present description, any concentration range, percentage range, ratio range, or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated. As used herein, "about" means.+-.10% of the indicated range, value, sequence, or structure, unless otherwise indicated. It should be understood that the terms "a" and "an" as used herein refer to "one or more" of the enumerated components unless otherwise indicated or dictated by its context. The use of the alternative (e.g., "or") should be understood to mean either one, both, or any combination thereof of the alternatives. As used herein, the terms "include" and "comprise" are used synonymously. In addition, it should be understood that the individual single chain polypeptides or immunoglobulin constructs derived from various combinations of the structures and substituents described herein are disclosed by the present application to the same extent as if each single chain polypeptide or heterodimer were set forth individually. Thus, selection of particular components to form individual single chain polypeptides or heterodimers is within the scope of the present disclosure

[0340] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0341] It is to be understood that the methods and compositions described herein are not limited to the particular methodology, protocols, cell lines, constructs, and reagents described herein and as such may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the methods and compositions described herein, which will be limited only by the appended claims.

[0342] All documents, or portions of documents, cited in the application including, but not limited to, patents, patent applications, articles, books, manuals, and treatises are hereby expressly incorporated by reference in their entirety for any purpose. All publications and patents mentioned herein are incorporated herein by reference in their entirety for the purpose of describing and disclosing, for example, the constructs and methodologies that are described in the publications, which might be used in connection with the methods, compositions and compounds described herein. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the inventors described herein are not entitled to antedate such disclosure by virtue of prior invention or for any other reason.

EXAMPLES

[0343] The following specific and non-limiting examples are to be construed as merely illustrative, and do not limit the present disclosure in any way whatsoever. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present disclosure to its fullest extent. All publications cited herein are hereby incorporated by reference in their entirety. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.

[0344] Exemplary bi-specific anti-CD3-CD19, anti-CD3-CDH3, anti-CD3-HER2, anti-CD3-HER3 and anti-CD3-EGFR antigen-binding constructs were made as described below. An exemplary schematic representation of these type of constructs is shown in FIGS. 1A-D. All formats are based on the heterodimeric Fc constructed by known mutations in the CH3 domain (Von Kreudenstein et al., MAbs. 2013 5(5):646-54). Exemplary constructs were conjugated to drugs to make ADCs using exemplary drugs DM1, DM4 and MMAE.

Example 1. Description, Expression and Purification of Bi-Specific Anti-CD19-CD3 Antigen-Binding Constructs Useful for ADCs in Dual scFv Format

[0345] Bispecific antibodies against CD3 and CD19 were designed, expressed and characterized as described in PCT/US2015/011664. Briefly, the genes encoding the antibody heavy and light chains were constructed via gene synthesis using codons optimized for human/mammalian expression. The scFv-Fc sequences were generated from a known anti-CD3 and CD19 scFv BiTE.TM. antibody (Kipriyanov et. al., 1998, Int. J Cancer: 77,763-772), anti-CD3 monoclonal antibody OKT3 (Drug Bank reference: DB00075). The dual scFv variants made are described in Table 1.

TABLE-US-00010 TABLE 1 Dual scFv variants Variant Chain A Chain B Fc 873 .alpha.CD19_HD37 scFv .alpha.CD3 Het Fc 1 (blinatumomab) scFv 875 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 1 1661 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2; Fc.gamma.R KO 2 1653 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2 (CDR C->S) 1662 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2; (CDR C->S) Fc.gamma.R KO 2 1660 .alpha.CD3_OKT3 scFv .alpha.CD19_HD37 scFv Het Fc 2 (VHVL linker) 1666 .alpha.CD3_OKT3 scFv .alpha.CD19_HD37 scFv Het Fc 2; (VHVL linker) Fc.gamma.R KO 2 1801 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2 (VLVH SS) N1 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2; (VLVH SS) Fc.gamma.R KO 2 6747 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2 (VLVH SS) (VLVH SS) 10149 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2; (VLVH SS) (VLVH SS) Fc.gamma.R KO 2 N3 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2 (VLVH SS) (CDR C->S) (VLVH SS) 10150 .alpha.CD19_HD37 scFv .alpha.CD3_OKT3 scFv Het Fc 2; (VLVH SS) (CDR C->S) (VLVH Fc.gamma.R KO 2 SS) 1380 .alpha.CD19_HD37 scFv .alpha.CD3_BiTE scFv Het Fc 2; Fc.gamma.R KO 1 N10 .alpha.CD19_HD37 scFv, .alpha.CD3_OKT3 scFv Het Fc 2 humanized (VLVH SS) (VLVH SS)

[0346] Het Fc 1=Chain A: L351Y_F405A_Y407V; Chain B: T366L_K392M_T394W (EU numbering system for IgG1 Fc) [0347] Het Fc 2=Chain A: T350V_L351Y_F405A_Y407V; Chain B: T350V_T366L_K392L_T394W [0348] Fc.gamma.R KO 1=Chain A: L234A_L235A; Chain B: L234A_L235A [0349] Fc.gamma.R KO 2=Chain A: D265S_L234A_L235A; Chain B: D265S_L234A_L235A [0350] .alpha.CD19_HD37 scFv--N- to C-terminal order of variable regions is VL/VH unless otherwise indicated [0351] .alpha.CD3_OKT3 scFv--N- to C-terminal order of variable regions is VL/VH unless otherwise indicated. The VLVH are connected by a (GGGGS).sub.3 linker. [0352] .alpha.CD3_BiTE scFv--N- to C-terminal order of variable regions is VH/VL and linker and composition is identical to blinatumomab. [0353] (VLVH SS) or (VHVL SS) indicates disulfide stabilized scFv utilizing the published positions VH 44 and VL 100, according to the Kabat numbering system, to introduce a disulphide link between the VH and VL of the scFv [Reiter et al., Nat. Biotechnol. 14:1239-1245 (1996)]. [0354] (CDR C->S)--indicates a mutation in the H3 CDR of OKT3 as referenced below [0355] (VHVL linker)--indicates VH and VL connected by the linker SSTGGGGSGGGGSGGGGSDI.

[0356] Fc numbering is according to EU index as in Kabat referring to the numbering of the EU antibody (Edelman et al., 1969, Proc Natl Acad Sci USA 63:78-85); Fab or variable domain numbering is according to Kabat (Kabat and Wu, 1991; Kabat et al, Sequences of proteins of immunological interest. 5th Edition--US Department of Health and Human Services, NIH publication no. 91-3242, p 647 (1991)).

[0357] The variants described in Table 1 include variant 875, a preliminary design, which was used as a starting point to generate antigen-binding constructs with improved yield and biophysical properties. The modifications include stabilization of the scFv by VLVH disulfide engineering and/or adding stabilizing CDR mutations. All variants include a heterodimeric Fc (Het Fc 1 or Het Fc 2) and can be expressed with or without mutations in the CH2 domain (Fc.gamma.R KO 1 or Fc.gamma.R KO 2) to abolish Fc effector activity. Variants including this modification to the Fc are referred to as having an Fc knockout or Fc KO.

[0358] Variants 875, 1661, 1653, 1662, 1660, 1666, 1801, and 1380 are initial designs of the CD3-CD19 antigen-binding constructs developed, while variants 6747, 10149, and 12043 exemplify designs that include modifications designed to further improve yield and biophysical properties of the CD3-CD19 antigen-binding constructs (see Example 3-4 for additional details). Variants N1, N3 and N10 have also been designed and the biophysical and functional characteristics of these variants can be predicted from the data provided herein.

[0359] The VHVL disulfide engineering strategy for both the CD3 and CD19 scFvs utilized the published positions VH 44 and VL 100, according to the Kabat numbering system, to introduce a disulphide link between the VH and VL of the scFv [Reiter et al., Nat. Biotechnol. 14:1239-1245 (1996)]. The mutation of C to S in the H3 CDR of .alpha.CD3 OKT3 scFv was generated as described in Kipryanov et al., in Protein Engineering 10: 445-453 (1997).

[0360] The final gene products were sub-cloned into the mammalian expression vector pTT5 (NRC-BRI, Canada) and expressed in CHO cells (Durocher, Y., Perret, S. & Kamen, A. High-level and high-throughput recombinant protein production by transient transfection of suspension-growing CHO cells. Nucleic acids research 30, E9 (2002)).

[0361] The CHO cells were transfected in exponential growth phase (1.5 to 2 million cells/mL) with aqueous 1 mg/mL 25 kDa polyethylenimine (PEI, Polysciences) at a EI:DNA ratio of 2.5:1. (Raymond C. et al. A simplified polyethylenimine-mediated transfection process for large-scale and high-throughput applications. Methods. 55(1):44-51 (2011)). In order to determine the optimal concentration range for forming heterodimers, the DNA was transfected in optimal DNA ratios of the heavy chain A (HC-A), and heavy chain B (HC-B) that allow for heterodimer formation (e.g. HC-A/HC-B/ratios=50:50%). Transfected cells were harvested after 5-6 days with the culture medium collected after centrifugation at 4000 rpm and clarified using a 0.45 .mu.m filter.

[0362] The clarified culture medium was loaded onto a MabSelect SuRe (GE Healthcare) protein-A column and washed with 10 column volumes of PBS buffer at pH 7.2. The antibody was eluted with 10 column volumes of citrate buffer at pH 3.6 with the pooled fractions containing the antibody neutralized with TRIS at pH 11. The protein was desalted using an Econo-Pac 10DG column (Bio-Rad).

[0363] In some cases, the protein was further purified by gel filtration, 3.5 mg of the antibody mixture was concentrated to 1.5 mL and loaded onto a Superdex 200 HiLoad 16/600 200 pg column (GE Healthcare) via an AKTA Express FPLC at a flow-rate of 1 mL/min. PBS buffer at pH 7.4 was used at a flow-rate of 1 mL/min. Fractions corresponding to the purified antibody were collected, concentrated to .about.1 mg/mL and stored at -80.degree. C.

[0364] An additional purification step using, protein L chromatography after protein a purification could be carried out by the method as follows. Capto L resin was equilibrated with PBS and the variant was added to the resin and incubated at RT for 30 min. The resin was washed with PBS, and bound protein was eluted with 0.5 ml 0.1 M Glycine, pH 3.

[0365] The purity and yield of the final product was estimated by LC/MS and UPLC-SEC as described in detail in PCT/US2015/011664. All variants were expressed and purified to >95% heterodimer purity without contaminating homodimers.

[0366] The clones that correspond to each bi-specific anti-CD3-CD19 antigen-binding construct are shown in Table XX (at the end of the Examples), and the corresponding sequence composition of each clone is shown in Table YY. The CDR sequences used in the variants are shown in Table S1.

TABLE-US-00011 TABLE S1 CDR sequences CD3 and CD19 antigen binding constructs Antigen binding constructs CDR sequence Wild-type OKT3 (CD3 binding) L1: SSVSY L2: DTS L3: QQWSSNP H1: GYTFTRYT H2: INPSRGYT H3: ARYYDDHYCLDY Stabilized VARIANT of OKT3 (CD3 binding) L1: SSVSY L2: DTS L3: QQWSSNP H1: GYTFTRYT H2: INPSRGYT H3: ARYYDDHYSLDY Humanized VARIANT of OKT3 (CD3 binding) short L1: SSVSY L2: DTS L3: QQWSSNP H1: GYTFTRYT H2: INPSRGYT H3: ARYYDDHYSLDY Humanized VARIANT of OKT3 (CD3 binding) long L1: SASSSVSYMN L2: DTSKLAS L3: QQWSSNPFT H1: GYTFTRYTMH H2: YINPSRGYTN H3: YYDDHYSLDY HD37 (CD19 binding) short L1: QSVDYDGDSYL L2: DAS L3: QQSTEDPWT H1: GYAFSSYW H2: IWPGDGDT H3: RETTTVGRYYYAMDY Humanized VARIANT of HD37 (CD19 binding) short L1: QSVDYEGDSYL L2: DAS L3: QQSTEDPWT H1: GYAFSSYW H2: IWPGDGDT H3: RETTTVGRYYYAMDY Humanized VARIANT of HD37 (CD19 binding) short L1: QSVDYSGDSYL L2: DAS L3: QQSTEDPWT H1: GYAFSSYW H2: IWPGDGDT H3: RETTTVGRYYYAMDY HD37 (CD19 binding) long L1: KASQSVDYDGDSYL L2: DASNLVS L3: QQSTEDPWT H1: GYAFSSYWMN H2: QIWPGDGDTN H3: RETTTVGRYYYAMDY Humanized VARIANT of HD37 (CD19 binding) long L1: RASQSVDYEGDSYL L2: DASNLVS L3: QQSTEDPWT H1: GYAFSSYWMN H2: QIWPGDGDTN H3: RETTTVGRYYYAMDY Humanized VARIANT of HD37 (CD19 binding) long L1: RASQSVDYSGDSYL L2: DASNLVS L3: QQSTEDPWT H1: GYAFSSYWMN H2: QIWPGDGDTN H3: RETTTVGRYYYAMDY

Example 2: Description, Expression and Purification of Exemplary Bi-Specific Antigen-Binding Anti-CD3-CD19 Constructs in a Hybrid Heterodimer Fc Format or in Full-Size Antibody Format

[0367] Additional bi-specific anti-CD3-CD19 antigen-binding constructs 1853, 6754, 10151, 6750, 6751, 6475, 6749, 10152, 10153, and 6518 were prepared as described in Example 1. These constructs are based on the same antigen-binding domains as variant 875 but have been engineered for improved yield and biophysical properties. The modifications include changing one or both scFvs to the equivalent Fab format and/or stabilization of the scFv by VL-VH disulfide engineering and stabilizing CDR mutations. The details of the variant compositions are shown in Table 2.

TABLE-US-00012 TABLE 2 Summary of Variants and Composition Variant # WT Fc (FcgR knock-out) * Chain 1 Chain 2 Dual scFv 875 (1661) .alpha.CD3_OKT3 scFv .alpha.CD19_HD37 scFv heterodimer 873 .alpha.CD3_blinatumomab scFv .alpha.CD19_HD37 scFv Fc variants 1653 .alpha.CD3_OKT3 scFv (CDR C->S) .alpha.CD19_HD37 scFv Hybrid 1853 (6754) .alpha.CD3_Teplizumab Fab .alpha.CD19_HD37 scFv heterodimer N5 (10151) .alpha.CD3_Teplizumab Fab .alpha.CD19_HD37 scFv (VHVL SS) Fc variants 6750 (6751) .alpha.CD3_OKT3 scFv .alpha.CD19_HD37 Fab 6475 (6749) .alpha.CD3_OKT3 scFv (CDR C->S) .alpha.CD19_HD37 Fab N7 (10152) .alpha.CD3_OKT3 scFv (VLVH SS) .alpha.CD19_HD37 Fab N11 (10153) .alpha.CD3_OKT3 scFv (CDR C->S) .alpha.CD19_HD37 Fab (VLVH SS) 6476 .alpha.CD3_blinatumomab scFv .alpha.CD3_HD37 Fab Full size mAb 6518 (N12) .alpha.CD3_Teplizumab Fab .alpha.CD19_HD37 Fab * All variants have the following CH3 mutations: Chain 1: T350V_L351Y_F405A_Y407V; Chain 2: T350V_T366L_K392L_T394W Variants in brackets refer to the equivalent Fc knockout variant that include the additional mutations D265S_L234A_L235A on both heavy chains. This abolishes binding of the Fc to Fc.gamma.Rs.

[0368] The anti-CD19 scFv and anti-CD3 scFv sequences were generated as described above. The anti-CD19 Fab (HD37 Fab) is a chimeric Fab using the HD37 VH and VL sequences fused to human IgG1 CH and CL sequences respectively. The scFv or VH-CH domains are fused to one chain of the heterodimeric Fc. The anti-CD3 Fab (tepizumab Fab) was generated from the known sequence of humanized OKT3 antibody teplizumab (Eli Lilly). The VH-CH domain was fused to one chain of the heterodimeric Fc.

[0369] The scFv disulfide engineering strategy (VHVL SS) for both the anti-CD3 and anti-CD19 scFvs utilized the published positions VH 44 and VL 100, according to the Kabat numbering system, to introduce a disulphide link between the VH and VL of the scFv [Reiter et al., Nat. Biotechnol. 14:1239-1245 (1996)].

[0370] The following variants contain a mutation to the anti-CD3 scFv to improve stability and yield, as reported previously [Kipriyanov et al., Prot. Eng. 10(4):445-453 (1997)]. v1653, v6475 and v10153 have an anti-CD3 (OKT3) with Cysteine to Serine mutation at position 100A of the VH CDR3.

[0371] Details of the cloning, expression and characterization of hybrid and full sized anti-CD3-CD19 antigen-binding constructs are provided in PCT/US2014/046436.

[0372] The clones that correspond to each bi-specific anti-CD3-CD19 and antigen-binding construct are shown in Table XX, and the corresponding sequence composition of each clone is shown in Table YY.

[0373] Controls

[0374] v891 has a polypeptide sequence that is identical to blinatumomab (BiTE.TM.) and includes an anti-CD3 scFv and anti-CD19 scFv (50 kDa).

[0375] Variant 4371 is a bivalent monospecific anti-CD19 antibody (used in Seattle Genetics' anti-CD19 antibody-drug conjugate known as SGN-19A denintuzumab mafodotin.)

[0376] In some experiments, polyclonal human IgG is used as a control and is referred to as v6249.

Example 3. Humanization and Stabilization of Anti-CD3 and Anti-CD19 Antibodies

[0377] The known murine and humanized anti-CD3 and CD19 antibodies BiTE.TM. antibody (Kipriyanov et. al., 1998, Int. J Cancer: 77,763-772), anti-CD3 monoclonal antibody OKT3 (Drug Bank reference: DB00075), anti-CD3 monoclonal antibody teplizumab (Drug Bank reference: DB00075) and anti-CD19 monoclonal antibody HD37 (Kipriyanov et. al., 1998, Int. J Cancer: 77,763-772; Pezzutto, A. et al., 1986, Leukocyte Typing II. Vol. 2. Springer-Verlag. HeidelberglNewYork. P. 391.) exhibit low production yield and biophysical stability.

[0378] To improve the yield and biophysical properties of the HD37 and OKT3 based antibodies we used a structure guided approach for humanization and stabilization. This approach is based on the humanization and stabilization method as described by Ewert et al., (Ewert et al., Methods 34 (2004) 184-199) and in addition includes detailed analysis of the VH/VL three dimensional structures to identifying potential VH/VL framework positions responsible for the low stability. Further, the framework and CDR sequences were analyzed for potential sites of post-translational modifications, including de-amidation, aspartate isomerization and protease cleavage.

[0379] The engineered humanized anti-CD3 and anti-CD19 VL and VH sequences and the sequence alignment to the known parental murine antibodies HD37 and OKT3 and the humanized teplizumab are shown in FIGS. 2 and 4 respectively. Critical positions identified by the structure guided humanization and stabilization approach are underlined and highlighted in bold in FIGS. 2 and 4. The engineered humanized sequences indicated hVH/hVL were used for construction of the bispecific variants as described in Example 4.

Example 4. Expression and Purification of Bi-Specific Anti-CD19-CD3 Antigen-Binding Constructs with Improved Yield and Biophysical Properties

[0380] Bispecific anti-CD3-CD19 antibodies designed for improved yield and biophysical stability were constructed as described in Table 3 and Example 3. Variant v10149 and v6751 are initial murine dual scFv heterodimer Fc and hybrid heterodimer Fc designs of the CD3-CD19 antigen-binding constructs (see Example 1 and Example 2 for further description). Variants v12043 and v15192-v15195 exemplify humanized designs that include variable domain framework and CDR modifications designed to further improve yield and biophysical properties of the CD3 and CD19 antigen-binding constructs.

[0381] The anti-CD19 murine HD37 scFv has been described in Example 1 and the Fab anti-CD19 murine HD37 is a chimeric Fab using the HD37 VH and VL sequences fused to human IgG1 CH and CL sequences respectively. The humanized HD37 Fab is a Fab composed of the humanized HD37 VH and VL sequences hVH2 and hVL2(D-E) (FIG. 2) fused to human IgG1 CH and CL sequences respectively. The humanized HD37 scFv is composed of the humanized HD37 VH and VL sequences hVH2 and hVL2(D-E) (FIG. 2) and has the identical VH/VL orientation and linker as described for v10149 above. The murine anti-CD3 scFv is identical to the scFv in the parental variant v875 (Table 1) and the humanized anti-CD3 scFvs were generated from the engineered VH and VL sequences as described in FIG. 4 and Table 3. [0382] (VLVH SS) indicates disulfide stabilized scFv utilizing the published positions VH 44 and VL 100, according to the Kabat numbering system, to introduce a disulphide link between the VH and VL of the scFv [Reiter et al., Nat. Biotechnol. 14:1239-1245 (1996)].

TABLE-US-00013 [0382] TABLE 3 Summary of Variants and Composition Bispecific Anti-CD19 chain on Anti-CD3 chain on variant # heavy chain A heavy chain B VH/VL mutations for improved stability v10149 murine HD37 scFv murine OKT3 scFv Original murine HD37 and OKT3 VH/VL sequences (VLVH SS) (VLVH SS) v12043 murine HD37 scFv murine OKT3 scFv Original murine OKT3 VH/VL sequences (VLVH SS) (VLVH SS) VHVL framework mutations for HD37 CDR mutation Asp->Glu at position 28 of HD37 VL v6751 murine HD37 Fab murine OKT3 scFv Original murine HD37 and OKT3 VH/VL sequences v15192 humanized HD37 Fab humanized OKT3 scFv VHVL framework mutations for HD37 and OKT3 (hVH1/hVL1) CDR mutation Cys->Ser at position 100A of OKT3 VH CDR mutation Asp->Glu at position 28 of HD37 VL v15193 humanized HD37 Fab humanized OKT3 scFv VHVL framework mutations for HD37 and OKT3 (hVH1/hVL2) CDR mutation Cys->Ser at position 100A of OKT3 VH CDR mutation Asp->Glu at position 28 of HD37 VL v15194 humanized HD37 Fab humanized OKT3 scFv VHVL framework mutations for HD37 and OKT3 (hVH2/hVL1) CDR mutation Cys->Ser at position 100A of OKT3 VH CDR mutation Asp->Glu at position 28 of HD37 VL v15195 humanized HD37 Fab humanized OKT3 scFv VHVL framework mutations for HD37 and OKT3 (hVH2/hVL2) CDR mutation Cys->Ser at position 100A of OKT3 VH CDR mutation Asp->Glu at position 28 of HD37 VL v 17119 humanized OKT3 scFv humanized HD37 Fab VHVL framework mutations for HD37 and OKT3 (hVH2/hVL2) CDR mutation Cys->Ser at position 100A of OKT3 VH CDR mutation Asp->Glu at position 28 of HD37 VL

[0383] The humanized Fab and scFv sequences are fused to the heterodimer Fc chains as described for the parental murine variants in Examples 1 and 2. All variants have the following CH.sub.3 mutations: Heavy chain A: T350V_L351Y_F405A_Y407V; Heavy chain B: T350V_T366L_K392L_T394W. The respective heavy chain CH3 mutations can either be on the anti-CD19 chain or the anti-CD3 chain. All variants further comprise the additional mutations D265S_L234A_L235A on both heavy chains to abolish binding of the Fc to Fc.gamma.Rs.

[0384] Fc numbering is according to EU index as in Kabat referring to the numbering of the EU antibody (Edelman et al., 1969, Proc Natl Acad Sci USA 63:78-85); Fab or variable domain numbering is according to Kabat (Kabat and Wu, 1991; Kabat et al, Sequences of proteins of immunological interest. 5th Edition--US Department of Health and Human Services, NIH publication no. 91-3242, p 647 (1991)).

[0385] The murine HD37 and OKT3 sequences were humanized and further modified for improved yield and stability by the following changes: i) the humanized anti-CD3 scFvs can utilize the published Cysteine to Serine mutation at position 100A of the VH CDR3 [Kipriyanov et al., Prot. Eng. 10(4):445-453 (1997)] and the variants v15192-v15195 and v17119 (Table 3) contain the Serine mutation for improved stability, ii) the sequence of the humanized anti-CD19 CDR was modified at position 28 to eliminate a potential Aspartate isomeration site that could impact antigen binding, iii) specific VHVL framework positions were identified to potentially impact stability and yield (Example 3); these positions are underlined and highlighted in bold in FIGS. 2 and 4.

[0386] The clones that correspond to each bi-specific anti-CD3-CD19 and antigen-binding construct are shown in Table XX, and the corresponding sequence composition of each clone is shown in Table YY.

[0387] Include Table

[0388] The bispecific antibodies against CD3 and CD19 were designed, expressed and characterized as described in PCT/US2015/011664 and in Examples 1 and 2.

[0389] The bispecifc antibodies were purified by Protein A affinity chromatography and subsequent gel filtration, as described in Example 1. FIG. 6 and Table 4 show the results of the preparative SEC purification and the final post purification yield of the initial parental murine variant v6751 and the engineered humanized variants.

[0390] The initial murine variant v6751 shows close to 50% high molecular aggregates after protein A purification and a low overall yield, while the engineered variants show minimal aggregates and 2-3-fold increased yield. The final post purification yield is comparable to positive control commercial antibodies.

TABLE-US-00014 TABLE 4 Production yield of humanized variants Purification Post pA/SEC yield Sample Name process (mg/L) v10149 pA/SEC 2.5 v12043 pA/SEC 5.2 v6751 pA/SEC 9.2 v15192 pA/SEC 25.2 v15193 pA/SEC 27.8 v15194 pA/SEC 29.6 v15195 pA/SEC 21.2

Example 5. Thermal Stability of Engineered Bi-Specific Anti-CD19-CD3 Antigen-Binding Constructs

[0391] The thermal stability of the stability engineered bispecific anti-CD19-CD3 constructs in comparison to the murine parental variants was assessed by differential scanning calorimetry (DSC).

[0392] All DSC experiments were carried out using a GE VP-Capillary instrument. The proteins were buffer-exchanged into PBS (pH 7.4) and diluted to 0.3 to 0.7 mg/mL with 0.137 mL loaded into the sample cell and measured with a scan rate of 1.degree. C./min from 20 to 100.degree. C. Data was analyzed using the Origin software (GE Healthcare) with the PBS buffer background subtracted.

[0393] Table 5 shows a list of the estimated melting temperatures (Tm) for the individual anti-CD3 and anti-CD19 Fab and scFvs of the parental murine vs. the stability engineered humanized constructs.

TABLE-US-00015 TABLE 5 A: Thermal stability of engineered anti-CD19 binding domains Anti-CD19 binding domain Tm (DSC) mHD37 scFv 53.degree. mHD37 Fab 65.degree. hHD37 Fab (hVH2/hVL2(D-E)) ~72.degree.(*) .sup. B: Thermal stability of engineered anti-CD3 binding domains Anti-CD3 binding domain Tm (DSC) OKT3 scFv .sup. 63.degree. Teplizumab Fab .sup. 66.degree. Teplizumab scFv ~62.degree.(*) hOKT3 scFv (hVH2/hVL2) ~72.degree.(*) (*)the DSC was measured on variants in IgG format; due to the overlap of CH2, Fab and scFv transitions with similar melting temperatures the specific Tm could only be estimated (see FIG. 7)

[0394] The anti-CD19 and anti-CD3 Fabs and scFvs were constructed as described above (Examples 1 and 4), expressed as bispecific or monospecific Fc constructs and the purified constructs were measured by DSC as described. FIG. 7 illustrates a representative DSC thermogram of selected engineered variants vs. the parental murine control. The melting transitions of the individual domains as summarized in Table 5 were estimated by comparison of the engineered vs. the parental murine DSC thermograms.

[0395] The results in Table 5 and FIG. 7 show that the humanized constructs with engineered variable domains have increased stability compared to their murine parental constructs. The final stabilized hybrid variants v15192-v15195 have thermal melting temperatures of over 72.degree. C., comparable to Fabs of commercial IgG antibodies.

[0396] As illustrated in Table 4 and 5, the structure guided stability engineering yields a significant improvement in expression and thermal stability. Further, comparison to the humanized Teplizumab shows that the improvement in yield and stability is independent of the sequence humanization, but is most likely due to specific changes to VH/VL positions that we have identified by our structure guided approach as critical for the Fab/scFv stability.

[0397] In conclusion, our structure guided humanization and stabilization approach has identified new humanized OKT3 and HD37 VH/VL sequences with significantly improved yield and stability. In difference to the known murine and humanized HD37 and OKT3 scFvs, which exhibit low expression and stability, our engineered variants show yield and stability comparable to commercial IgG and thus allow the development as therapeutic antibodies.

Example 6. Antigen Binding of Engineered Bi-Specific Anti-CD19-CD3 Antigen-Binding Constructs

[0398] To determine whether the engineered bispecific constructs v15192-v15195 bind to CD19 and CD3 antigens equivalent to the parental murine construct v6751, the binding affinity to CD19 and CD3 was measured by SPR and whole cell FACS as described below.

[0399] All SPR binding experiments were carried out using a BioRad ProteOn XPR36 instrument at 25.degree. C. with 10 mM HEPES, 150 mM NaCl, 3.4 mM EDTA, and 0.05% Tween 20 at pH 7.4. Recombinant CD3epsilon/delta Fc fusion protein (Sino Biological; http://www.sinobiological.com/CD3D-CD3-Delta-Protein-g-10182.html) was captured on anti-Fc capture sensorchips. Purified antibodies were indirectly captured on the sensorchip by binding the recombinant CD3 fusion protein when injected at 25 .mu.L/min for 240s (resulting in approx. 500 RUs) following a buffer injection to establish a stable baseline. Resultant K.sub.D values were determined from binding isotherms using the Equilibrium Fit model with reported values as the mean of three independent runs.

[0400] Table 6 summarizes the results of the SPR binding of the engineered humanized bispecific constructs v15192-v15195. All engineered constructs bind to CD19 and CD3 antigens equivalent to the parental murine construct v6751. The stability engineered humanized constructs have equivalent binding to CD3 antigen compared to the parental v6751.

TABLE-US-00016 TABLE 6 SPR binding of engineered anti-CD19- CD3 variants to recombinant CD3 Sample capture (RU) KD (M) Rmax (RU) v6751 959.93 1.41E-07 178.72 v15193 988.48 3.70E-07 174.01 v15194 975.92 3.49E-07 179.12 v15195 1032.89 4.27E-07 192.69

Example 7: Whole Cell Binding to CD19+ Raji Tumor Cells and CD3+ Jurkat T Cells and Human PBMCs

[0401] The ability of the humanized bispecific anti-CD19-CD3 constructs to bind to CD3- and CD19-expressing cells was assessed via whole cell FACS binding analysis as described previously (PCT/US2015/011664). The binding to CD19+ Raji B cells (ATCC: CCL-86; Pulvertaft, Lancet 1964) and CD3+ Jurkat cells (Weiss, J Immunol 1984) and the apparent binding affinities of variant v15195 are shown in FIGS. 8A and B.

[0402] The bispecific anti-CD19-CD3 constructs (exemplified by v15195 in FIG. 8) bound human Raji CD19+ NHL B cells with high affinity (apparent affinity of 2.4 nM) and human CD3+ Jurkat T cells with low affinity (apparent affinity of 44.7 nM).

[0403] The ability of bispecific T cell engagers to crosslink T cells and target B cells is a prerequisite for activity. Therefore, in addition to assessing binding to isolated B and T cell lines as shown in FIGS. 8A and B we tested the ability of the bispecific anti-CD19-CD3 constructs to crosslink autologous B and T cells in human PBMC. Freshly isolated human and PBMCs were incubated with v15195 on ice and the percentage of B cell:T cell doublets were analyzed by FACS to determine the concentration dependent ability of crosslinking B and T cells. The percent T:B doublets were defined as FSC-W-high cells within the CD20+ SSClow population. Greater than 75% of the identified doublets were CD4+ or CD8+, suggesting that they had formed doublets with T cells.

[0404] As illustrated in FIG. 8C, the analysis of B:T cell doublets in human PBMC demonstrated that v15195 crosslinks B and T cells in human PBMC in a concentration dependent manner and at concentrations below 1 nM.

[0405] Together, this data shows that the bispecific anti-CD19-CD3 constructs preferentially binds B cell and crosslinks B and T cells at concentrations below 2 nM, while binding to isolated T cells at significantly lower concentrations of above 40 nM. This preferential binding of B cells and crosslinking of B and T cells at low concentrations, while only binding isolated T cells at low concentrations, allows for development of bispecific drug-conjugates that will preferentially bind B cells and activate T cells without impacting isolated T cells.

Example 8: Drug Conjugation of Bi-Specific Anti-CD19-CD3 Antigen-Binding Constructs

[0406] A schematic of exemplary anti-CD3-CD19 antigen-binding construct drug conjugate is shown in FIG. 1. Anti-CD3-CD19 antigen-binding constructs were conjugated to either DM1 using the non-cleavable linker SMCC or to DM4 using the cleavable linker SPBD as described below. Variants were conjugated to either DM1 or DM4 using a one-step procedure. The starting protein sample was first exchanged into a buffer composed of 50 mM potassium phosphate pH 6.5, 50 mM NaCl and 2 mM EDTA using a PD-10 column, and adjusted to a protein concentration of 2-10 mg/ml. A 10 mM solution of SMCC-DM1 (Levena Biopharma US, San Diego, Calif.) or SPBD-DM4 (Levena Biopharma US, San Diego, Calif.) dissolved in dimethylacetamide (DMA) was then added to 7.5 molar equivalents of the protein sample. DMA was further added to a final concentration of 10% v/v and the sample was mixed briefly. The reaction mixture was incubated at 25.degree. C. overnight with mixing. The product was then exchanged into a buffer composed of 20 mM sodium succinate pH 5.0 using a PD-10 column, and the protein concentration and drug-to-antibody ratio (DAR) were calculated based on the absorbance at 252 and 280 nm. The buffer was adjusted to a final composition of 20 mM sodium succinate, 6% w/v trehalose and 0.02% w/v polysorbate 20, pH 5.0. High performance liquid chromatography-size exclusion chromatography (HPLC-SEC) was performed to determine the purity of the ADC, using the Tosoh G3000-SWXL column (7.8 mm.times.30 cm), in 100 mM sodium phosphate, 300 mM sodium chloride, pH 7.0, at a flow rate of 1 ml/min.

[0407] SMCC-DM1 conjugates of v12043, v6754, 6751, 15195 and 4372 had a yield of over 70%, a purity of >85% and a drug/antibody ratio (DAR) of 2.2-3.5 as summarized in Table 7. The SPBD-DM4 conjugate of v12043 had a yield of 70% and a purity of 82%.

TABLE-US-00017 TABLE 7 Conjugation of bispecific anti-CD3-CD19 variants Variants Conjugate DAR % purity Yield 12043 SMCC-DM1 2.2 91 71 12043 SPBD-DM4 2.8 82 70 6754 SMCC-DM1 2.5 85 75 6751 SMCC-DM1 3.5 97 72 4372 SMCC-DM1 3.5 90 78 15195 SMCC-DM1 3.5 97 72

[0408] FIG. 9 shows an exemplary UPLC-SEC profile of v12043-SMCC-DM1 after conjugation. Other variants behaved similarly.

Example 9. Impact of Bispecific Format on In Vitro Activity of Anti-CD3-CD19 Antigen-Binding Drug-Conjugates Against B and T Tumor Cell Lines

[0409] To test the cytotoxicity and potency of anti-CD3-CD19 conjugates on target B cells and T cells, selected bispecific anti-CD3-CD19 conjugates with identical CDRs, but differing antigen binding format of scFv or Fab were tested in a growth inhibition assay using B and T tumor cell lines. The selected variants in FIG. 10 and Table 8 have previously been shown to have similar binding affinities to CD19 and CD3. The affinity of all selected variants to CD19 is .about.2 nM, the affinity to CD3epsilon is .about.40 nM.

[0410] The extent of cytotoxicity was measured in cell cultures of CD19+ Raji or Ramos (ATCC: CRL-1596; Klein, Intervirology 1975) non-Hodgkin lymphoma (NHL) target B cell lines and CD3+ Jurkat T cell line in comparison to non-specific IgG SMCC-DM1 conjugate (v6249) and monospecific anti-CD19 antibody huBU12 conjugated to SMCC-DM1 as positive control (v4371). The monospecific anti-CD19 antibody huBU12 is currently being evaluated as a MC-MMAF drug conjugate (denintuzumab mafodotin) in Phase I and Phase II clinical trials in NHL and B-ALL (Gerber, Blood 2009; Albertson TM, Proceeding: AACR Annual Meeting 2014). Potential off-target cytotoxicity of the SMCC-DM1 conjugates was measured against the target cell line, K562 (ATCC: CCL-243) which does not express CD19 or CD3. The selected antibodies were diluted in media and added to the target Raji, Ramos, Jurkat or K562 cells in triplicate and incubated for 24 hr. Cells were washed, media replaced and cell survival was evaluated after a 3 day incubation at 37.degree. C. Cell viability was measured using Sulforhodamine B with absorbance read at 510 and 540 nm following standard procedures. Data was normalized to untreated control and analysis was performed in GraphPad prism.

[0411] All anti-CD3-CD19 conjugates showed no off-target activity against the cell line K562 which does not express CD19 or CD3, similar to the non-specific IgG-SMCC-DM1 control v6249 (data not shown).

TABLE-US-00018 TABLE 8 Cytotoxicity of selected anti-CD3-CD19 variants on B and T cells Ramos Jurkat Therapeutic IC50 IC50 Window (IC50 Rank Variant Format Linker-toxin (nM) (nM) Jurkat/Ramos) 1 6751 .alpha.CD3 (scFv)- SMCC-DM1 0.4562 23.39 51.3 .alpha.CD19 (Fab) 2 12043 .alpha.CD3 (scFv)- SMCC-DM1 2.982 26.55 8.9 .alpha.CD19 (scFv) (batch 2) 3 12043 .alpha.CD3 (scFv)- SPBD-DM4 0.2885 2.121 7.4 .alpha.CD19 (scFv) 4 12043 .alpha.CD3 (scFv)- SMCC-DM1 0.5399 2.496 4.6 .alpha.CD19 (scFv) (batch 1) 5 4371 .alpha.CD19 control SMCC-DM1 6.025 24.33 4.0 6 6249 Non-specific SMCC-DM1 37.14 134.7 3.6 IgG control 7 6754 .alpha.CD3 (Fab)- SMCC-DM1 3.974 0.1891 0.0 .alpha.CD19 (scFv) (batch 1) 8 6754 .alpha.CD3 (Fab)- SMCC-DM1 6.251 0.1672 0.0 .alpha.CD19 (scFv) (batch 2)

[0412] The growth inhibition results, as illustrated in FIG. 10 and Table 8, show unexpectedly, that the cytotoxic activity on target B cells of all monovalent anti-CD19 bispecific antibodies is comparable to or better than the bivalent monospecific anti-CD19-SMCC-DM1 control v4371. The conjugated variant 6751 has a potency of 0.45 nM on Ramos B cells and the bivalent monospecific positive control v4371 conjugate has a potency of .about.6 nM.

[0413] The growth inhibition results suggest in addition an unexpected difference between the different hybrid and dual scFv anti-CD3-CD19-MCC-DM1 conjugates. Variant 6751-MCC-DM1 (with the anti-CD3 in scFv format and the anti-CD19 in Fab format) is highly active towards B cells with EC50 of 0.45 nM, while having very low activity on Jurkat T cells, similar to the non-specific controls v4371 and v6249. In contrast, the variant v6754 (with the anti-CD3 in Fab format and the anti-CD19 in scFv format) has similar potency on target B cells and T cells. A potential therapeutic window of killing target B cell without impacting T cells was calculated as shown in Table 8. The data suggest a therapeutic window and killing of target B cell without impacting the T cells for the variant 6751 and 12043, but not 6754.

[0414] These results show that unexpectedly, a bispecific T cell engager drug conjugate can be developed to preferentially bind and kill target B cells, while not impacting the T cells. As result of to the preferential binding and activity, the bispecific T cell engager drug conjugate has the potential to have a dual mechanism of action of: i) T cell redirected B cell killing and ii) B cell killing through internalization of the conjugated toxin payload.

[0415] Further, the data suggests the preferential behaviour is dependent on one or all of the following characteristics of the bispecific: i) monospecific targeting of the CD3 antigen, ii) low affinity binding to the CD3 antigen, iii) format and geometry of the bispecific. In conclusion, the results allow the identification of the bispecific format (including Fab vs scFv and hybrid vs. dual scFv or full size Ig bispecific and Ig isotype and hinge) and CD3e affinity as the critical parameters that have to be optimized for the development of bispecifc CD3 T cell engager drug conjugates.

[0416] This conclusion and the ranking of variants with different format is confirmed in activity assays with primary T cells as described in Example 10 and also in internalization assays with tumor and T cell lines (Example 21).

Example 10. In Vitro Efficacy of Exemplary Anti-CD3-CD19 Antigen-Binding Construct Drug-Conjugates in Primary Human Blood Samples

[0417] To further test the preferential killing of target B cells without affecting T cells and T cell activity, the selected variants were tested in primary blood cultures with allogeneic Ramos and Raji lymphoma cell lines. This assay reflects the cytotoxic activity of the anti-CD3-CD19 conjugates towards the allogeneic target B cells mediated by the T cell redirected activity of the bispecific, and also the conjugated drug delivered by internalization of the antigen-binding construct by the target B cells. To measure the effect of the conjugates on the T cell population the T cell activity, activation and proliferation were analyzed. As relevant markers for total T cell counts CD4 and CD8 have been measured whereas T cell activation of the CD8 and CD4 T cells was measured by the established early and late T cell activation markers CD69 and CD25, respectively.

[0418] In addition, the T cell exhaustion marker PD-1 was measured to evaluate the potential effect on T cell inhibition and exhaustion. PD-1 (Programmed cell Death protein 1) functions as an immune checkpoint and plays an important role in down regulating the immune system by preventing the activation of T-cells and promoting T cell apoptosis, while reducing apoptosis in regulatory T cells (suppressor T cells) (Francisco L M, Sage P T, Sharpe A H (July 2010) Immunological Reviews 236: 219-42)

[0419] Human blood (120-140 mL) for individual studies was collected from donors and PBMC were freshly isolated. PBMCs were further processed to derive the subpopulations without autologous B cells (PBMC-B). Resting PBMCs were used as effector cells and Raji or Ramos human B cells as target cells and the ratio of T cells to allogeneic Raji or Ramos B cells was adjusted to an E:T ratio of 5:1. B cell and T cell populations, at day 0, were determined by FACS. Exclusive B cell markers included CD19 and CD20. T cell populations were measured by CD3, CD4 and CD8 and T cell activation and potential exhaustion was measured by CD69, CD25 and PD1, respectively as described above. Quadruplicate wells were plated for each control and experimental condition and co-cultures were incubated in 5% CO2, 37.degree. C. and stopped at 72 hours. T and B cells were assessed for their respective proportions in the culture by FACS. The collected culture cells were stained for CD45, CD20 and 7-AAD FACS detection. FACS analysis was carried out by InCyte/FlowJo as follows: A Guava 8HT flow cytometer was used for analysis of cell subpopulations. Between 5,000 events for FSC/SSC and compensation wells, and 30,000 events for experimental wells were analyzed by cytometry. A threshold was set to skip debris and RBCs. All B cells were confirmed to be double positive for CD19 and CD20 at Day 0, which allowed for monitoring of CD20 as appropriate B cell marker. In a control experiment Raji and Ramos cell cultures without PBMC were incubated with the variants analyzed for B cell cytotoxicity after 72 hrs.

[0420] FIGS. 11 and 12 show the results of an n=2 repeat with two individual PBMC donors and allogeneic Raji B cells. The non-conjugated variant v12043 has a potency of <0.05 nM on the Raji target B cells for both donors and induces T cell proliferation and activation with similar potency. The non-conjugated variant v12043 is able to deplete .about.50% of target B cells by the T cell redirected mechanism. In contrast, the drug conjugates show an equivalent T cell mediated B cell depletion at concentrations below 0.5 nM, but in addition are able to further deplete the target Raji B cells at concentrations above 0.5 nM. Unexpectedly, the conjugates show only at the highest concentration of 50 nM an impact on the T cell proliferation, but do not have an impact at lower concentrations. This is in line with the data presented in FIG. 10 and Table 8.

[0421] FIGS. 13 and 14 show the results of a separate repeat experiment of the hybrid variants v6751 and v6754 with Raji and Ramos target B cells using fresh PBMCs from the same donor as in FIG. 12. The activity of the anti-CD3-CD19 variants is compared to the positive controls blinatumomab and the anti-CD19 monospecific conjugate. The results in FIGS. 13 and 14 show a similar additional activity of the conjugates on the target B cell depletion at higher concentrations compared to the non-conjugated v6751 and v6754. As suggested by the growth inhibition assay in FIG. 10, the assay confirms the unexpected difference of v6751 and v6754 conjugates on the T cell, with v6754 conjugate having no impact on the T cells while mediating potent killing of the target B cells.

[0422] In addition to the B cell depletion and T cell counts measured in the previous experiments (FIGS. 11 and 12), the up-regulation of PD-1 was measured. PD-1 plays an important role in down regulating the immune system by preventing the activation of T-cells and promoting T cell apoptosis (Francisco L M, Sage P T, Sharpe A H (July 2010) Immunological Reviews 236: 219-42). PD-1 up-regulation has been speculated to be a mechanism of resistance to T cell redirected therapies [Junttila et al., Cancer Res (2014) 5561-71; Kohnke, 2015]. As shown in FIG. 14 with PBMC and allogeneic Raji cells all variants, including the positive control blinatumomab induced up-regulation of PD-1 in >80% of T cells and no significant B cell depletion of the non-conjugated variants. The conjugated variants were able to deplete the Raji B cells at higher concentrations, but not the non-conjugated variants. In addition, the anti-CD3-CD19 conjugates in comparison to the non-conjugated comparators show a lower % of PD-1 expressing T cells at higher concentrations. T

[0423] The clinical and preclinical experience of Blinatumomab indicate that the T cell redirected response is highly donor dependent and can be limited by mechanisms of T cell immunosuppression (Kohnke, 2015). As illustrated above all tested bispecifics (including blinatumomab) induced up-regulation of PD1 and in some donors the unconjugated bispecific were ineffective in depleting the target B cells. In contrast, the conjugated bispecific T cell engager showed activity in these cultures, suggesting that the dual mechanism of action can potentially overcome limited efficacy in patients with high T cell immunosuppression.

Example 11. Cytotoxicity of Bispecific Anti-CD19-CD3-SMCC-DM1 Drug Conjugates Against ALL, NHL Tumor Cell Lines Grown in Culture without T Effector Cells

[0424] To test the cytotoxicity and potency of the humanized bispecific anti-CD3-CD19 variants with improved biophysical properties (see Example 3-5), selected variants were conjugated to SMCC-DM1 as described in Example 8. All SMCC-DM1 conjugates of v6751, v15192, v15193, v15194, v15195 had comparable yield of over 70%, purity of >90% and a drug/antibody ratio (DAR) of 3.1-3.5.

[0425] The extent of cytotoxicity was measured in cell cultures of different CD19+ non-Hodgkin lymphoma (NHL) and acute lymphocytic leukemia (ALL) tumor B cell lines in comparison to non-specific IgG SMCC-DM1 conjugate (Isotype DM1) and monospecific anti-CD19 antibody huBU12 conjugated to auristatin as positive control. The monospecific anti-CD19 antibody huBU12 is currently being evaluated as a MC-MMAF drug conjugate (denintuzumab mafodotin) in Phase I and Phase II clinical trials in NHL and B-ALL (Gerber, Blood 2009; Albertson.TM., Proceeding: AACR Annual Meeting 2014).

[0426] The impact on T cells is tested on CD3+ Jurkat T cells. Potential off-target cytotoxicity of the SMCC-DM1 conjugates was measured against the target cell line, K562 which does not express CD19 or CD3. The experiment was conducted as described in detail in Example 9.

[0427] FIG. 15 illustrates the results for a selected subset of target cell lines and Table 9 summarizes the results in comparison to the anti-CD19 antibody positive control.

TABLE-US-00019 TABLE 9 Cytotoxicity of MCC-DM1 drug conjugates against ALL, NHL tumor cell lines grown in culture without T cells v15195-MCC- huBU12-MCC- DM1 DM1 Target cell line (IC50 nM) (IC50 nM) ALL (CD19.sup.+, CD3.sup.-) Nalm-6 (ATCC: CRL-3273) 0.7 ~5 * RS4; 11 (ATCC: CRL-1873) <5 * <5 * DLBCL (CD19.sup.+, CD3.sup.-) SUDHL-4 (ATCC: CRL-2957) 1.2 4.7 SUDHL-6 (ATCC: CRL-2959) 1.8 <5 * Burkitt (CD19.sup.+, CD3.sup.-) Raji ** 1.5 6.9 Ramos ** 0.4 6.0 Daudi (ATCC: CCL-213) 2.1 ~5 * T-cell leukemia (CD19.sup.-, CD3.sup.+) Jurkat 24.3 23.4 AML (CD19.sup.-, CD3.sup.-) K562 (ATCC: CCL-243) Greater Greater than 50 nM than 50 nM

[0428] For the results indicated with * only a 5 point concentration curve was measured and the Kd could not be fitted with confidence. The results of e.g. <5 indicates that at the concentration of 5 nM over 50% of cells were depleted. The results indicated with ** refer to data collected for the murine v6751-SMCC-DM1 conjugate.

[0429] As shown in Table 9, the bispecific anti-CD3-CD19 drug conjugates show potent killing of NHL and ALL tumor B cells lines while not significantly impacting the growth of the Jurkat T cells. All anti-CD3-CD19 conjugates showed no off-target activity against the cell line K562, which does not express CD19 or CD3, similar to the non-specific IgG-SMCC-DM1 control (data not shown).

[0430] In addition, the potency was comparable or greater than the positive control huBU12-MCC-DM1 and v15195-MCC-DM1 exhibited a wide range of target cell cytotoxic killing across human cancer cell lines.

Example 12. Cytotoxicity of Bispecific Unconjugated Anti-CD19-CD3 and Bispecific Anti-CD19-CD3-SMCC-DM1 Drug Conjugates Against Tumor Cell Lines Grown in Culture with T Cells

[0431] The target B cell cytotoxic activity of the SMCC-DM1 conjugated and unconjugated variant v15195 was further evaluated in comparison to the approved therapeutic antibody Blinatumomab. The bispecific variant v15195 was specifically chosen because of the over 100 fold lower T cell redirected potency compared to Blinatumomab. This lower T cell mediated potency is sufficient to mediate target B cell killing in vitro and in vivo, while resulting in lower T cell activation and proliferation compared to Blinatumomab at 1000 fold lower concentration (see Example 14). Importantly, the lower potency yields compatible potencies for the T cell redirected and DM1 mediated cytotoxicity and enables a dual mechanism of action.

[0432] The cytotoxic activity of the bispecific anti-CD3-CD19 conjugates was measured in comparison to the non-conjugated anti-CD3-CD19 variant and the positive control Blinatumomab.TM. (blinatumomab, BiTE.TM.). To measure the effect of the conjugates on the T cell population the T cell activity, activation and proliferation were further analyzed as described in Example 13. The assay was performed with n=4 primary blood donors and the experimental set-up was conducted as described above in Examples 11-14.

[0433] As illustrated in FIG. 16, comparison of the activity of the unconjugated v15195 and the DM1 conjugated v15195 confirms that the cytotoxic activity of the anti-CD3-CD19 conjugates towards the allogeneic target B cells can be mediated by the T cell redirected activity of the bispecific, but also by the conjugated drug delivered by internalization of the antigen-binding construct by the target B cells.

[0434] Further, the results show the benefit of a dual mechanism as the T cell mediated activity of both the unconjugated v15195 and the positive control Blinatumomab at efficacious concentrations is highly donor dependent and not sufficient to kill >90% of the target B cells in this assay.

Example 13. Cytotoxicity of Bispecific Anti-CD19-CD3-SMCC-DM1 Drug Conjugate Against Tumor Cell Lines Grown in Culture with T Cells

[0435] To further test the activity of the bispecific anti-CD19-CD3-SMCC-DM1 drug conjugates, the extent of cytotoxicity was measured in co-cultures of different CD19+ non-Hodgkin lymphoma (NHL) or acute lymphocytic leukemia (ALL) tumor B cell lines and primary T cells.

[0436] The variant v15195-MCC-DM1 was tested in primary blood cultures with allogeneic NHL or ALL cell line. The experimental set-up was conducted as described above in Example 12.

[0437] FIG. 17 shows potent killing of different NHL and ALL target B cell lines by the bispecific anti-CD19-CD3-SMCC-DM1 drug conjugate and confirms preferential killing of target B cells without impacting T cells. The T cell counts are not impacted up to the highest tested concentration of 50 nM (data not shown).

Example 14. T Cell Activation and Proliferation Effects of Bispecific Anti-CD19-CD3-SMCC-DM1 Drug Conjugate in Comparison to Blinatumomab and OKT3 Antibodies

[0438] The clinical dosing of the commercial therapeutic antibody Blinatumomab is limited by toxicities that are thought to be T cell mediated and associated with the extent of T cell proliferation and activation (Chatenoud, 1986; Abramowicz, 1989; Goebeler, 2011; Bargou, 2008; Topp, 2011; Klinger, 2010; International Patent Publication No. WO2011051307A1; Goebeler M E J Clin Oncol 2016; Topp, Lancet Oncol 2015)

[0439] To evaluate the potential therapeutic index of v15195, the ability of v15195 to induce T cell activation and proliferation was assessed in co-cultures of Raji cancer B cells and human PBMC and compared to the in vitro activity of Blinatumomab at a concentration equivalent to the clinically tolerated exposure. (The maximum tolerated doe (MTD) of Blinatumomab in Phase 1 r/r-NHL trial is 60 .mu.g/m2/day (Goebeler M E J Clin Oncol 2016); for comparison to a safe dose and exposure of Blinatumomab, a concentration of 0.05 nM Blinatumomab or equivalent to the exposure at the dose of 40 .mu.g/m2/day was chosen).

[0440] The co-culture experiment was conducted as follows: On Day 0, blood was collected from each of 4 donors and PBMCs were freshly isolated. PBMCs were further processed to derive the subpopulation of PBMC without B cells (PBMC-B). Resting PBMC-B were used as effector cells and Raji human B cells as target cells and the ratio of T cells to allogeneic Raji cells was adjusted to an E:T ratio of 5:1. The mixtures were incubated together with the antibody constructs for 3 days, after which the collected primary cells were stained for CD4, CD8, CD69, CD25 FACS detection. FACS analysis of the different populations was carried out by InCyte/FlowJo as follows: Between 5,000 event for FSC/SSC and compensation wells, and 30,000 events for experimental wells were analyzed by cytometry. A threshold was set to skip debris and RBCs.

[0441] FIG. 18A illustrates the results from n=4 donors after 72 h incubation for the CD8+ T cell populations. The analysis shows the total CD8+ T cell counts, which is an indirect measure of the induced T cell proliferation and also the extend of T cell activation, measured by the early and late T cell activation markers CD69 and CD25 (see Example 10), respectively.

[0442] The results show that at efficacious concentrations of 100-1000 fold above the in vitro EC.sub.50 (see FIG. 16 and Example 12), v15195 induced lower T cell proliferation and activation than clinically tolerated concentrations of Blinatumomab.

[0443] In addition to the FACS based analysis of induced T cell proliferation and activation, the v15195 induced T cell proliferation was evaluated in a thymidine cell proliferation assay in PBMC cultures, as depicted in FIG. 18B. The thymidine based assay presents a different measure of T cell proliferation and activation as it is a measure of the total induced proliferation potential in the PBMC culture. The thymidine based analysis provides a complementary measures to the FACS based method described above. In the assay illustrated in FIG. 18B, v6751, the original parental murine variant to v15195 was used (see Table 3).

[0444] The thymidine cell proliferation assay in PBMC cultures was conducted as follows: On Day 1, blood was collected from each of 3 donors and PBMCs were freshly isolated. The test items were prepared for a final concentration of 0.3 and 100 nM and combined with the PBMCs, plated at 250,000 cells/well. The mixtures were incubated for 3 days, after which tritiated thymidine was added to the cell containing wells for a final of 0.5 .mu.Ci thymidine/well; the plates were incubated for an additional 18 hours, after which the plates were frozen. Total incubation time was 4 days. The plates were filtered and counted (CPMs) using a .beta.-counter. From the averages, a Stimulation Index (SI) was calculated as follows and the data was tabulated: average CPM of test item/average CPM of media only. The average E:T ratio in human PBMC collected from healthy donors was .about.10:1 CD3+ T cells to CD19+ B cells.

[0445] As illustrated in FIG. 18B, the comparison to Blinatumomab and OKT3 demonstrated lower total cell proliferation in cultures of v6751 even at 1000 fold higher concentrations. This suggests that the bispecific CD19-CD3 drug conjugate does not impact T cell at to the highest evaluated concentration and further that the therapeutic index is potentially higher than Blinatumomab.

Example 15: Target B Cell-Dependence of T Cell Activation by Bi-Specific Heterodimer Variants in Human PBMC

[0446] The dependence of T-cell activation by the exemplary anti-CD19-CD3-SMCC-DM1 bi-specific variant v15195 on target B cells was determined in human PBMCs. The experiment was carried out as described below.

[0447] Human blood (120-140 mL) was collected from donors and PBMC were freshly isolated from donors. PBMCs were further processed to derive the subpopulation of PBMC without B cells (PBMC-B). Quadruplicate wells were plated for each control and experimental condition and PBMC cultures were incubated in 5% CO2, 37.degree. C. and stopped at 72 hours. T cell populations were assessed by FACS. The cell pellets were resuspended in various antibody cocktails for flow cytometry analysis. A Guava 8HT flow cytometer was used for analysis of cell subpopulations. As negative controls an anti-CD19 bivalent monospecific antibody (huB12; see Example 11) and untreated cultures were used. The results are shown in FIG. 19.

[0448] The results indicate that v15195 does not activate T cells in cultures of PBMC lacking B cells, but activates T cells in presence of target B cells. Variant v15195 shows strictly target dependent T cell activation.

Example 16. T Cell Activation, Proliferation and Cytokine Release of Bispecific Anti-CD19-CD3-SMCC-DM1 Drug Conjugate v15195, in Comparison to Unconjugated v15195

[0449] The ability of the bispecific SMCC-DM1 conjugated and un-conjugated constructs to induce T cell proliferation and activation was assessed in two different assays as described below. The assay was performed with n=4 primary blood donors and the experimental set-up was conducted identical to Examples 12 and 14.

[0450] FACS Analysis of T Cell Proliferation/Activation in Raji/PBMC-B Cultures:

[0451] On Day 0, blood was collected from each of 4 donors and PBMCs were freshly isolated. PBMCs were further processed to derive the subpopulation of PBMC without B cells (PBMC-B). Resting PBMC-B were used as effector cells and Raji human B cells as target cells and the ratio of T cells to allogeneic Raji cells was adjusted to an E:T ratio of 5:1. The mixtures were incubated together with the antibody constructs for 3 days, after which the collected primary cells were stained for CD4, CD8, CD69, CD25 FACS detection.

[0452] Cytokine Analysis of Raji/PBMC-B Co-Culture Supernatant:

[0453] Raji co-culture experiments were set-up as described above and levels of IFN-.gamma., IL-6 and IL-10 were assessed by luminex after 3 days of incubation.

[0454] The results of the FACS based analysis of T cell proliferation and activation in PBMC-B/Raji co-cultures is shown in FIG. 20A. Cytokine production is presented in FIG. 20B.

[0455] The results in FIGS. 20A and 20B illustrate that conjugation of v15195 with SMCC-DM1 enhanced T cell proliferation and activation at low concentrations, compared to unconjugated v15195. In addition, SMCC-DM1 conjugated v15195 enhanced production of the pro-inflammatory cytokines, IFN-.gamma. and IL-6 compared to unconjugated v15195. While SMCC-DM1 conjugated v15195 only induced a modest increase in the anti-inflammatory cytokine IL-10, the unconjugated form caused a dose-dependent increase.

[0456] The difference in T cell activation and particularly cytokine profile is an unexpected and relevant result, since for example IL10 release has been associated with T cell suppressive mechanisms (e.g. regulatory T cell expansion) which limit the efficacy of T cell engagers. The bispecific T cell engager drug conjugate could therefore potentially be less susceptible to T cell suppressive mechanisms. In addition the increased release of the pro-inflammatory cytokine INF.gamma., which is a key regulator for T cell and macrophage activation, has the potential to synergize and enhance the T cell redirected activity.

[0457] This difference in T cell activation and cytokine release is dose dependent and correlates with the dose response of DM1 mediated target cell depletion, which suggests that the effect is mediated by the activity of the drug conjugate. As previously reported, DM1 and DM1-ADCs can mediate immunogenic cell death and are highly synergistic with immunomodulatory agents, like anti-PD1 and anti-CTLA4 (Mueller et al., Science Transl Med 2015). The results suggest that addition of a toxin like DM1 has the potential to improve the efficacy of the anti-CD19-CD3 bispecific, by inducing immunogenic/pro-inflammatory cell death (Mueller et al., Science Transl Med 2015).

Example 17. Cytotoxicity of Bispecific Anti-CD19-CD3 Conjugated to MMAE Against Tumor Cell Lines Grown in Culture with T Cells

[0458] To further test the preferential killing of target B cells without affecting T cells, v15193 was conjugated to the toxin MMAE using a cleavable linker (maleimidocaproyl-valine-citrulline-p-aminobenzyloxycarbonyl (mc-vc-PABC)). The cytotoxic activity of the bispecific anti-CD3-CD19 conjugates was measured in comparison to the non-conjugated anti-CD3-CD19 variant in primary blood cultures with allogeneic Ramos lymphoma cell lines. Possible cytotoxicity towards T cells as a result of drug conjugation using a cleavable linker was also assessed.

[0459] To prepare the antibody drug conjugate, the antibody was first bound to a "Lock-Release"resin (ADCR01, ADC Biotechnology Ltd.), a proprietary resin for immobilization of antibodies for conjugation, at 1 mg/100 .mu.L resin loading. Antibody disulfide binds were then reduced by addition of a solution of tris(carboxyethyl)phosphine (TCEP) in PBS (pH7.4) with 2 mM EDTA to the bound antibody at 6 molar equivalents of TCEP to bound antibody and incubation of the mixture at 20.degree. C. for 120 minutes with continuous mixing. Excess TCEP was removed by washing (.times.3) with PBS (pH7.4).

[0460] For conjugation, a solution of mc-vc-PABC-MMAE (ADC Biotechnology Ltd., see structure below) (10 mM stock in dimethyl acetamide (DMA)) equivalent to 6 molar equivalents of linker-toxin to bound antibody was first prepared in PBS containing 5% (v/v) DMA. This linker-toxin solution was added to the bound antibody and the mixture incubated at 20.degree. C. for 60 minutes with continuous shaking.

##STR00020##

[0461] Following conjugation, excess linker-toxin solution was removed by centrifugation of the resin at 14800 rpm for 2 minutes. The resin was then washed three times with PBS containing 5% (v/v) DMA to remove any residual linker-toxin, followed by three washes with PBS (pH 7.4) to remove any remaining DMA co-solvent. The antibody-drug conjugate was released from the resin by incubating the resin for 15 minutes in Release buffer (ADC Biotechnology Ltd.) followed by centrifugation at 14800 rpm for 2 minutes. The filtrate was then desalted by G25 gel permeation chromatography (GE Healthcare Illustra.TM. NAP.TM.-5 column) into a buffer containing 10 mM sodium acetate, 9% sucrose (pH 5.0), followed by filtration through a sterile 0.22 .mu.m PES membrane.

[0462] Purity of the final antibody-drug conjugate was assessed by high performance liquid chromatography-size exclusion chromatography (HPLC-SEC) on a TSKgel G3000SWXL 7.8 mm.times.30 cm, 5 .mu.m column (TOSOH Bioscience LLC) in 10% IPA, 0.2M potassium phosphate, 0.25M potassium chloride, pH 6.95 at a flow rate of 0.5 mL/min. The drug-to-antibody ratio (DAR) of the antibody-drug conjugate was determined using hydrophobic interaction chromatography (HIC) HPLC on a Butyl-NPR 4.6 mm.times.3.5 cm, 2.5 .mu.m column (TOSOH Bioscience LLC) run at 0.8 mL/min with a 12-minute linear gradient of A--1.5M (NH.sub.4).sub.2SO.sub.4, 25 mM NaPi, pH 6.95 and B--25 mM NaPi, pH 6.95, 25% IPA.

[0463] The final yield of the v15193-mc-vc-PABC-MMAE conjugate was 39%, with a purity of >98% and an average DAR of 3.7.

[0464] To measure the effect of the conjugate on the T cell population, the T cell activity, activation and proliferation were further analyzed as described in Example 14. The assay was performed with n=1 primary blood donors and the experimental set-up was conducted as described above in Example 12.

[0465] As illustrated in FIG. 21, comparison of the activity of the unconjugated and the MMAE conjugated bispecific variant (v15193 vs. v15193-mc-vc-PABC-MMAE ("v15193-vc-MMAE")) confirms that the cytotoxic activity of the anti-CD3-CD19 conjugate towards the allogeneic target B cells can be mediated by the T cell redirected activity of the bispecific, but also by the conjugated drug delivered by internalization of the antigen-binding construct by the target B cells.

[0466] Further, the results show unexpectedly that v15193-mc-vc-PABC-MMAE had little to no effect on T cell counts suggesting that bispecific T cell engager drug conjugates can be developed both with non-cleavable linkers (as described above), and with cleavable linkers such as mc-Val-Cit-PABC.

Example 18. In Vivo Response to Exemplary Anti-CD3-CD19 Antigen-Binding Construct Drug-Conjugates in Humanized Hu(CD34+)NSG Mice

[0467] To further evaluate the impact of anti-CD3-CD19 conjugates on the T cell population and activity, selected variants were analyzed in an in vivo study in humanized mice. The in vivo B cell depletion and activation and redistribution of autologous T cells was measured in humanized (CD34+) NSG mice (E:T .about.1:5) after a single dose IV injection of v12043 SMCC-DM1 and SPDB-DM4 conjugates in comparison to the non-conjugated v12043.

[0468] For humanization of mice, 2 week-old NSG (NOD scid gamma, NOD.Cg-Prkdcscid Il2rgtmlWjl/SzJ) mice were injected with human (CD34+) HSC from human fetal liver (Jackson Laboratory. Humanized (CD34+) NSG mice develop human T cell and B cell linages within 12 weeks. Average T cell to B cell ratio in humanized (CD34+) NSG is .about.1:5 to 1:1.

[0469] Humanized (hCD34+) NSG were dosed with 1 intravenous (IV) bolus injection at day 0 (at 0.3 and 0.1 mg/kg doses) and the autologous circulating B and T cell populations were analysed at 48 h post injection and at day 5 upon termination, similar to previously described (PCT/US2015/011664). The T cell and B cell populations were analyzed by FACS. The specific B and T cell markers analyzed were human CD45, CD20, CD4, CD8 and CD69, as described above in Example 10.

[0470] The in vivo serum exposure of the variants, as shown in Table 5, was estimated from previous data in NSG mice (see PCT/US2015/011664).

TABLE-US-00020 TABLE 10 Estimated serum exposure for v12043 Serum conc. 0.3 mg/kg 0.5 h >30 nM 24 h 10 nM 48 h 6 nM 72 h 3 nM 120 h 1 nM

[0471] FIG. 22 shows the impact on the B and T cell counts in circulation after single dose injection of v12043 and the v12043 SMCC-DM1 and SPBD-DM4 conjugates. All variants and dose levels were effective in depletion the circulating B cells (CD20+ B cells) and no significant difference on T cell counts and T cell activation, as measured by CD69 expression on CD4 and CD8 positive T cells, was observed between the groups.

[0472] The estimation of the serum exposure in Table 10 suggests a Cmax of close to 50 nM, which was the maximum concentration used in the in vitro assay in primary blood cultures. The in vivo single dose study confirms that at even at the highest dose of 0.3 mg/kg and a Cmax close to 50 nM, the anti-CD3-CD19 conjugates have no negative impact on T cells. Further, the conjugates do not reduce the T cell activation and T cell redirected activity on the B cells.

Example 19. In Vivo Response of Anti-CD3-CD19 Antigen-Binding Construct Drug-Conjugates in Hu(CD34+)NSG Mice

[0473] To further evaluate the impact of anti-CD3-CD19 conjugates on the T cells and T cell activation at higher doses and maximum exposures of over 300 nM, v15195-MCC-DM1 was analyzed in an in vivo study in humanized mice. The in vivo activation and redistribution of autologous T cells was measured in humanized (CD34+) NSG-SGM3 (NSG strain: NOD.Cg-Prkdc.sup.scid Il2rg.sup.tm1Wjl Tg(CMV-IL3,CSF2,KITLG)1Eav/MloySzJ; Jackson laboratory) mice (E:T<1:5) after a single dose IV injection at 9 mg/kg, 3 mg/kg, 1 mg/kg, and 0.3 mg/kg. As control, a variant with only the anti-CD19 Fab connected to the heterodimeric Fc, but lacking the anti-CD3 scFv (v15760) was used. The monospecific anti-CD19 control variant v15760 was conjugated to SMCC-DM1 as described in Example 8, with a purity of >90% and DAR of 3.5.

[0474] Humanized (CD34+) mice present a good model system to evaluate human T cell activation, redistribution and expansion in a mouse model, whereas the proliferation and maturation of human B cells is partially deficient in these models (Ito et al., Cellular & Molecular Immunology 2012; 9: 208-214; Brehm et al., Curr Opin Endocrinol Diabetes Obes. 2010; 17(2): 120-125). We therefore don't expect to see a significant effect of the conjugated drug DM1 on the human B cells, whereas the human T cell activation and proliferation is well established in these models and the main aim of this study is to assess the impact of the anti-CD3-CD19 bispecific drug conjugate on the T cell activation and expansion.

[0475] The study was conducted as previously described in Example 18 and PCT/US2015/011664. Briefly, humanized (hCD34+) NSG-SGM3 mice were purchased from Jackson laboratory. The bispecific anti-CD3-CD19 ADC, v15195-MCC-DM1 was dosed with 1 intravenous (IV) bolus injection at day 0 (at 0.3, 1, 3 and 9 mg/kg doses) and the autologous circulating B and T cell populations in peripheral blood and isolated spleen were analysed at day 8 upon termination, similar to previously described (PCT/US2015/011664). The T cell and B cell populations were analyzed by FACS. The specific B and T cell markers analyzed were human CD45, CD20, CD4, CD8 and CD69, as described above in Example 10 and 18.

[0476] FIG. 23 shows the total CD3+ T cell counts in peripheral blood and isolated spleen at day 8 post injection of the single dose. The results show that only at the selected highest dose of 9 mg/kg a significant effect on the T cells in circulation and in the spleen was observed. No impact on T cell counts and activation was observed at 3 mg/kg and lower doses.

[0477] The in vivo serum exposure of v15195, as shown in Table 11, was estimated from previous data of the unconjugated variants in NSG mice (PCT/US2015/011664).

[0478] The estimated serum exposure in Table 11 suggests that the bispecific T cell engager -MCC-DM1 conjugates can be dosed up to at least 3 mg/kg and an associated Cmax of >300 nM, without long term impact on the autologous T cells. This is an exposure and Cmax similar to other SMCC-DM1 antibody frug conjugates in clinical development and thus allows the development of T cell engager drug conjugates at dose levels that are standard for other -MCC-DM1 ADCs in development (Jumbe at al., J Pharmacokinet Pharmacodyn (2010) 37:221-242; Lu et al., Cancer Chemother Pharmacol (2014) 74:399-410).

TABLE-US-00021 TABLE 11 Estimated serum exposure Serum conc. 9 mg/kg 3 mg/kg 1 mg/kg 0.3 mg/kg 0.5 h >900 nM >300 nM >30 nM >30 nM 24 h 300 nM 100 nM 10 nM 10 nM 48 h 180 nM 60 nM 6 nM 6 nM 72 h 90 nM 30 nM 3 nM 3 nM 120 h 30 nM 10 nM 1 nM 1 nM

Example 20. Expression and Purification of Bi-Specific Anti-Tumor-CD3 Antigen-Binding Constructs for Solid Tumor Indications

[0479] Bispecific antibodies against CD3 and CDH3, HER2, HER3 or EGFR were designed, expressed and characterized as described in PCT/US2015/011664. Briefly, the genes encoding the antibody heavy and light chains were constructed via gene synthesis using codons optimized for human/mammalian expression. The scFv-Fc sequences were generated from a known anti-CD3 scFv BiTE.TM. antibody (Kipriyanov et. al., 1998, Int. J Cancer: 77,763-772) and anti-CD3 monoclonal antibody OKT3 (Drug Bank reference: DB00075). The CDH3 Fab sequences were generated from a known anti-CDH3 monoclonal antibody (PCT/JP2009/007333). The HER2 Fab sequences were generated from trastuzumab (PCT/US1998/026266, Baselga J et al., 1998, Cancer Res: 58,2825-31) and the HER3 Fab sequences were generated from a known anti-HER3 monoclonal antibody (PCT/EP2010/070062, Mirschberger C, et al., 2013, Cancer Res: 73, 5183-94). EGFR sequences were generated from cetuximab (PCT/US1996/009847, Prewett M et al., 1996, J Immunother Emphasis Tumor Immunol: 19,419-27). The Fab-scFv variants made are described in Table 12.

[0480] The humanized anti-CD3 OKT3 scFv was generated identical to anti-CD3 scFv of the murine OKT3 variants v875 or the humanized OKT3 variant v15195, described above. The anti-CD3 BiTEx-IC2 scFv was generated from the VH and VH sequences as described in (US 20110275787 A1), which is cross-reactive with non-chimpanzee primate CD3. The humanized OTK3 scFv or the BiTEx-IC2 scFv were fused to one chain of the heterodimeric Fc. The anti-CDH3 monoclonal, Clone #6 Fab is a chimeric Fab using the murine Clone #6 VH and VL sequences fused to human IgG1 CH and CL sequences respectively. The anti-HER2 monoclonal Fab consists of the humanized VH and VL sequences of trastuzumab fused to human IgG1 CH and CL sequences, respectively. The Fab of the anti-HER3 monoclonal is a fusion of the humanized VH and VL sequences of lumretuzumab (PCT/EP2010/070062; Mirschberger C., et al., 2013, Cancer Res., 73:5183-94) to human IgG1 CH and CL sequences, respectively. The anti-EGFR monoclonal, cetuximab Fab is a chimeric Fab using the murine cetuximab VH and VL sequences fused to human IgG1 CH and CL sequences, respectively. In all cases, the VH-CH domains of the antibodies are fused to the second chain of the heterodimeric Fc.

TABLE-US-00022 TABLE 12 Summary of Variants and Composition Bispecific variant # Anti-tumor antigen chain Anti-CD3 chain v13831 .alpha.CDH3_Clone #6 Fab BiTEx-I2C scFv (VL/VH) v13792 .alpha.HER2_(trastuzumab) Fab BiTEx-I2C scFv (VL/VH) v13790 .alpha.HER3_Mab205 Fab BiTEx-I2C scFv (VL/VH) v16371 .alpha.EGFR_(cetuximab) Fab BiTEx-I2C scFv (VL/VH) All variants have the following CH3 mutations: Heavy chain A: T350V_L351Y_F405A_Y407V; Heavy chain B: T350V_T366L_K392L_T394W. Chain A or B can be either on the anti-CD3 or the anti-tumor antigen chain.

[0481] Fc numbering is according to EU index as in Kabat referring to the numbering of the EU antibody (Edelman et al., 1969, Proc Natl Acad Sci USA 63:78-85); Fab or variable domain numbering is according to Kabat (Kabat and Wu, 1991; Kabat et al, Sequences of proteins of immunological interest. 5th Edition--US Department of Health and Human Services, NIH publication no. 91-3242, p 647 (1991)).

[0482] The clones that correspond to each bi-specific anti-tumor-CD3 and antigen-binding construct are shown in Table XX, and the corresponding sequence composition of each clone is shown in Table YY.

[0483] The bispecific antibodies against CD3 and CDH3, HER2, HER3 or EGFR were designed, expressed and characterized as described in PCT/US2015/011664 and in Examples 1 and 2.

[0484] The bispecifc antibodies were purified by Protein A affinity chromatography and subsequent gel filtration, as described in Example 1.

Example 21. In Vitro Internalization of Bispecific Anti-Tumor-CD3 pHAb-Conjugates on Solid Tumor Cell Lines and Jurkat T Cell Line

[0485] Conjugation of a pHAb dye to antibodies is a method used to assess internalization of a given antibody by a cell. The dyes only fluoresce under low pH conditions such as those found the the endosome/lysosome, indicating internalization of the antibody. Exemplary bispecific anti-tumor-CD3 antigen-binding construct pHAb conjugates were made as follows. Variants were conjugated to pHAb Amine Reactive Dye as per the manufacture's protocol for in-solution antibody conjugation (Promega).

[0486] The starting protein sample was first exchanged in 10 mM sodium bicarbonate buffer (pH 8.5) using a desalting column. A 10 mg/ml solution of pHAb Amine Reactive Dye dissolved in a 1:1 DMSO-water mix was then added at a 20 molar excess to the antibody sample. The reaction mixture was incubated for 60 minutes with mixing. Unreacted dye was removed using a desalting column. The antibody concentration and DAR were calculated after measuring absorbance at 280 nm and 532 nm. High performance liquid chromatography-size exclusion chromatography (HPLC-SEC) was performed to determine the purity of the conjugates, using the Superdex 200 column (8.6 5.times.150 mm), in D-PBS+0.01% Polysorbate 20, at a flow rate of 0.25 ml/min.

[0487] pHAb conjugates of v13831, v16371 had a yield of over 60%, a purity by HPLC-SEC of >90% and a drug/antibody ratio (DAR) of 1.5-3.3.

[0488] The extent of internalization was measured in several tumor cell lines, SKOV3 (ATCC: HTB 77), A431 (ATCC: CRL-1555), HCT-116 (ATCC: CCL-247) and JIMT1(AddexBio #C0006005), in comparison to non-specific IgG pHAb conjugate (v15195) and an anti-CD3 pHAb conjugated mAb, v2171 (UCHT1, Beverley PC and Callard R E., 1981, Eur J Immunol., 11: 329-34; PCT/US1993/007832). The anti-RSV antibody, Synagis (PCT/US1991/002668), was used as a negative control. The impact on T cells is tested on CD3+ Jurkat T cell. The selected antibodies were diluted in media and added to the target cells in triplicate and incubated for 1 hr. Cells were washed, media replaced and antibody internalization was evaluated using ImageExpress following standard procedures. Data was normalized to untreated control and analysis was performed in GraphPad prism.

[0489] The internalization results, as illustrated in FIG. 24 and Table 13 show that pHAb conjugated v13831 and v16371 are rapidly internalized by SKOV3, A431, HCT-116 and JIMT1 cells, with Kd values in the nM range and internalization of the variants is dependent upon expression of the target antigen. Unexpectedly, these variants are poorly internalized by the Jurkat T cell line. Furthermore, the bivalent anti-CD3 pHAb conjugated mAb (v2171, UCHT1) was internalized by Jurkat cells at close to 2 nM, while the bispecifics based on two different anti-CD3 scFvs, which target slightly different CD3e epitopes (OKT3 and xBiTE), showed low internalization into Jurkat T cells (72 nM and >100 nM, respectively).

[0490] Therefore, the data suggests that the bispecific tumor-CD3 antibodies are preferentially internalized by tumor cells compared to CD3.sup.+ cells making it less likely that ADC versions of these bispecifics would exhibit toxicity towards the T cells being engaged by the CD3 arm.

TABLE-US-00023 TABLE 13 Differential internalization of pHAb conjugated bispecific tumor-CD3 variants by tumor cell lines EC50 (nM) v13831 v16371 v15195 v2171 bispecific bispecific bispecific .alpha.CD3 control Target cell line (.alpha.CDH3-BiTEx) (.alpha.EGFR-BiTEx) (.alpha.CD19-hOKT3) (hUCHT1) Solid Tumor HCT-116 3.92 >100 (EGFR.sup.-/CDH3.sup.+) A431(EGFR.sup.+/CDH3.sup.+) 4.15 2.46 JIMT1 <10* 2.90 (EGFR.sup.+/CDH3.sup.+) SKOV3 >100 2.05 (EGFR.sup.+/CDH3.sup.-) T-cell leukemia Jurkat (CD19.sup.-, CD3.sup.+) >100 >100 71.95 13.2 *Estimated EC50; accurate fitting of the parameters was not possible due to increased background levels at antibody concentrations greater than 100 nM (see FIG. 24 for comparison).

[0491] The results illustrate further, that the concept of a bispecific T cell engager ADC with dual functionality, as established in Examples 1-14 for CD19-CD3 bispecifics can be expanded to different solid tumor antigens and solid tumor targeting bispecific T cell engagers. The preferential tumor targeting without impacting the T cells is not specific to the CD19 antigen, and the concept for the design of bispecific T cell engager ADCs, as described in Example 9, can be transferred to different tumor targets. In addition, the low impact on T cells is not specific to a particular epitope on CD3e, but likely rather dependent on the format and geometry of the bispecific, as described in Example 9.

Example 22. Cytotoxicity of Bispecific Anti-Tumor-CD3-SMCC-DM1 Drug Conjugates Against Breast, Ovarian Tumor Cell Lines Grown in Culture without T Cells

[0492] To test the cytotoxicity and potency of the bispecific anti-tumor-CD3 variants, selected variants were conjugated to SMCC-DM1 as described in Example 8. All SMCC-DM1 conjugates of v13831, v13792 and v13790 had comparable yield of over 70%, purity of >90% and a drug/antibody ratio (DAR) of 3.1-3.5.

[0493] The extent of cytotoxicity was measured in cell cultures of different breast (MCF7 (ATCC: HTB-22) and JIMT1) and ovarian (SKOV3) tumor cell lines in comparison to non-specific IgG SMCC-DM1 conjugate (Isotype DM1). The impact on T cells is tested on CD3+ Jurkat T cells. The experiment was conducted as described in detail in Example 9.

[0494] FIG. 25 illustrates the results for a selected subset of target cell lines and Table 14 summarizes the results in comparison to the IgG-DM1 control.

[0495] The results of the cytotoxicity study, as illustrated in FIG. 25 and Table 14 show that the DM1 conjugated variants, v13831, v13792, and v13790, exhibit potent killing of breast and ovarian tumor cell lines but do not significantly impact the growth of the Jurkat T cells. This preferential killing of tumor cell lines is similar to the preferential internalization of the variants by tumor cell lines presented in Example 22. The non-specific variant, v6249-SMCC-DM1, does not exhibit any significant killing of tumor cell lines or Jurkat T cells until concentrations greater than 100 nM are used. This highlights the role target specificity plays in the activity of the anti-tumor-CD3 bispecific antibodies.

TABLE-US-00024 TABLE 14 Cytotoxicity of SMCC-DM1 drug conjugates against breast, ovarian tumor cell lines grown in culture without T cells EC50 (nM) v13831-SMCC- v13792-SMCC- v13790-SMCC- v6249-SMCC- DM1 DM1 DM1 DM1 Target cell line (.alpha.CDH3-BiTEx) (.alpha.HER2-BiTEx) (.alpha.HER3-BiTEx) (hIgG) Breast cancer MCF7 3.1 12.6 3.6 50.2 JIMT1 1.4 3.4 -- 34.1 Ovarian cancer SKOV3 -- 0.1 -- 21.5 T-cell leukemia (CD19.sup.-, CD3.sup.+) Jurkat 12.2 16.7 15.3 53.7

[0496] Therefore, the data suggests that, like the CD19-CD3 bispecifics, the bispecific anti-tumor-CD3 drug conjugates preferentially kill tumor cells compared to CD3.sup.+ cells. This further supports the conclusion that bispecific ADCs would would possess potent anti-tumor activity while exhibiting little to no toxicity towards the T cells being engaged by the CD3 arm.

Example 23. Cytotoxicity of Unconjugated and SMCC-DM1 Conjugated Anti-Tumor-CD3 Bispecifics Against Tumor Cell Lines Grown in Culture with T Cells

[0497] To further test the preferential killing of target tumor cells without effecting T cells and T cell activity, selected variants were tested in primary blood cultures with allogeneic JIMT1 cell line. The cytotoxic activity of the bispecific anti-CD3-tumor conjugates was measured in comparison to the non-conjugated anti-CD3-tumor variant. To measure the effect of the conjugates on the T cell population the T cell activity, activation and proliferation were further analyzed as described in Example 23. The assay was performed with n=1 primary blood donors and the experimental set-up was conducted as described above in Example 5 with minor modifications described below.

[0498] Specifically, on day 0 JIMT1 cells were first labeled with CellTracer violet (a live/dead stain). Following the labeling of the JIMT1 target cells, PBMCs were isolated for use as effector cells. Rested PBMCs were mixed with the labeled JIMT1 cells such that the ratio of T cells to allogeneic JIMT1 cells was adjusted to an E:T ratio of 2:1. The mixtures were incubated together with the antibody constructs for 4 days, after which the JIMT1 cells were collected and viability was assessed though FACS analysis of CTV levels. This was carried out by InCyte/FlowJo as follows: Between 5,000 event for FSC/SSC and compensation wells, and 30,000 events for experimental wells were analyzed by cytometry. A threshold was set to skip debris and RBCs.

[0499] As illustrated in FIG. 26, both the unconjugated and SMCC-DM1 conjugated forms of the variants, v13831 and v13792 exhibited potent cytotoxic activity towards JIMT1 cells. The results show that this cytotoxic activity towards the allogeneic target tumor cells can be mediated by the T cell redirected activity of the bispecific. Interestingly, SMCC-DM1 conjugation of variants v13831 and v13792 enhanced tumor cell killing compared to the unconjugated versions of these variants at concentrations greater than 0.05 nM. This is likely a result of drug delivery to the target tumor cells through internalization of the antigen-binding construct.

[0500] Further, the results show the benefit of a dual mechanism as the T cell mediated activity of the unconjugated variants is highly donor dependent and not sufficient to kill >90% of the target tumor cells in this assay.

Example 24. T Cell Proliferation and Activation of Bispecific Anti-Tumor-CD3-SMCC-DM1 Drug Conjugates Compared to Unconjugated Anti-Tumor-CD3 Bispecifics

[0501] The ability of the SMCC-DM1 conjugated bispecifics, anti-CDH3-CD3 and anti-HER2-CD3, and their parent unconjugated constructs to induce T cell activation and proliferation was assessed as described below.

[0502] On Day 0, blood was collected from each of 4 donors and PBMCs were freshly isolated. Resting PBMC were used as effector cells and JIMT1 cells as target cells and the ratio of T cells to allogeneic JIMT1 cells was adjusted to an E:T ratio of 2:1. The mixtures were incubated together with the antibody constructs for 4 days, after which the collected primary cells were stained for CD4, CD8, CD69, and CD25. FACS analysis of the different populations was carried out by InCyte/FlowJo as follows: Between 5,000 event for FSC/SSC and compensation wells, and 30,000 events for experimental wells were analyzed by cytometry. A threshold was set to skip debris and RBCs.

[0503] The results of the FACS based analysis of T cell proliferation and activation in PBMC/JIMT1 co-cultures is shown in FIG. 27. The results illustrate that at efficacious concentrations, the SMCC-DM1 conjugated variants induce a modest increase in the total CD8.sup.+ and CD4.sup.+ T cell populations compared to the unconjugated parent variants. Total CD8.sup.+ and CD4.sup.+ T cell counts is an indirect measure of induced T cell proliferation suggesting that conjugation of a toxin to an anti-tumor-CD3 bispecific can enhance T cell proliferation induced by the unconjugated bispecific. Similarly, a modest increase in CD25.sup.+ and CD69.sup.+ (early and late stage T cell activation markers, respectively) T cells is also observed in when the co-cultures are stimulated with the SMCC-DM1 conjugated variants compared to the unconjugated variants, suggesting increased activation of the T cells.

Example 25. T Cell Proliferation and Activation of Bispecific Anti-Tumor-CD3-SMCC-DM1 Drug Conjugates Compared to Unconjugated Anti-Tumor-CD3 Bispecifics--Range of High, Intermediate and Low Effector to Target Ratios

[0504] To further delineate the role of each mechanism of action of the bispecific anti-tumor-CD3-SMCC-DM1 drug conjugates, T cell redirected killing and killing through internalization of the conjugated toxin payload, the cytotoxic activity of the anti-CD3-CDH3 conjugate, v13831-SMCC-DM1 was measured in comparison to its non-conjugated parent v13831. The variants were tested in primary blood cultures with allogeneic JIMT1 cell line at three different E:T ratios. The assay was performed with n=1 primary blood donors.

[0505] on day 0 JIMT1 cells were first labeled with CellTracer violet (a live/dead stain). Following the labeling of the JIMT1 target cells, PBMCs were isolated for use as effector cells. Rested PBMCs were mixed with the labeled JIMT1 cells such that the ratio of T cells to allogeneic JIMT1 cells was adjusted to an E:T ratios of 5:1, 1:5 and 1:50. The mixtures were incubated together with the antibody constructs for 4 days, after which the JIMT1 cells were collected and viability was assessed though FACS analysis of CTV levels. This was carried out by InCyte/FlowJo as follows: Between 5,000 event for FSC/SSC and compensation wells, and 30,000 events for experimental wells were analyzed by cytometry. A threshold was set to skip debris and RBCs.

[0506] The results presented in FIG. 28, suggest that at higher E:T ratios (eg. 5:1), the cytotoxic activity of the bispecific anti-CDH3-CD3 antibody, v13831 towards the allogeneic target tumor cells can be mediated by the T cell redirected activity of the bispecific. Thus, as the number of T effector cells decrease, as is the case in the 1:5 and 1:50 E:T ratios, tumor cell killing is nearly lost. Unlike the unconjugated variant, v13831-SMCC-DM1 was effective even at the lower E:T ratios (1:5 and 1:50) as a result of its second mode of action, drug delivery to the target tumor cells through internalization of the antigen-binding construct. Therefore, the data further supports the benefit of a dual mechanism of action as the T cell mediated activity of the unconjugated variants is highly donor dependent and not sufficient to kill >90% of the target tumor cells in this assay, especially when T effector cell concentrations are low. This may be of particular importance in indications where T cell infiltration of tumors is low.

TABLE-US-00025 TABLE XX H1 (Heavy H2 (Heavy L1 (Light L3 (Light Variant Chain 1 Chain 2 Chain 1 Chain 2 Number Clone No.) Clone No.) Clone No.) Clone No.) 873 1064 1065 875 1064 1067 1661 2183 2176 1653 1842 2167 1662 2183 2177 1660 2174 2175 1666 2184 2185 1801 1842 2228 6747 5243 2227 10149 6692 6689 10150 6692 6690 1380 1844 1890 12043 7239 6689 1853 2304 2175 6754 5239 2185 2309 10151 5239 6691 2309 6750 5241 5238 2310 6751 5242 2176 2310 6475 2305 2171 2310 6749 5242 2177 2310 10152 5242 6689 2310 10153 5242 6690 2310 6476 2305 2170 2310 6518 2304 2304 2309 2309 891 1109 4372 3344 3345 3346 3346 15192 9288 9284 9289 15193 9288 9285 9289 15194 9288 9286 9289 15195 9288 9287 9289 17119 11176 11177 11175 17118 11178 11179 11175 13831 8074 3320 8071 13792 1015 3320 -2 13790 3320 3299 16371 3537 3320 3357 873 1064 1065 17119 11176 11177 11175 17118 11178 11179 11175

TABLE-US-00026 TABLE YY SEQ ID Descrip- No. Clone tion Sequence 1. 2176 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY CLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 2. 2176 Full CAGATCGTCCTGACACAGAGCCCAGCTATCATGTCAGCAAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CCAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCCTCTGGAGTGCCTGCTCACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTGCAGCTGCAGCAGTCCGGAGCAGAGCTGGCT- CGACCAGGAGCTAGTGTGAAAATGTCCTGTAAGGC AAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGGCAGGGACTGGAATGGA- TCGGGTACATTAATCCTAGCCGAGGATACACAAACTAC AACCAGAAGTTTAAAGACAAGGCCACTCTGACCACAGATAAGAGCTCCTCTACCGCTTATATGCAGCTGAG- TTCACTGACATCTGAGGACAGTGCAGTGTACTATTGCGC CAGGTACTATGACGATCACTACTGTCTGGATTATTGGGGCCAGGGGACTACCCTGACAGTGAGCTCCGCAG- CCGAACCTAAATCTAGTGACAAGACTCATACCTGCCCCC CTTGTCCAGCACCAGAGGCTGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTG- ATGATCTCCCGGACACCTGAAGTCACTTGCGTGGTCGTG AGCGTGTCTCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCCACATATC GCGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGC- AACAAGGCACTGCCTGCCCCAATCGAGAAGACAATTA GCAAAGCAAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAA- AACCAGGTCAGTCTGCTGTGTCTGGTGAAGGGCTTCTA TCCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTG- TCCTGGACTCAGATGGGAGCTTCTTTCTGTATAGTAAAC TGACCGTGGACAAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCAC- AATCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGC AAG 3. 2176 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRG- SGSGTSYSLTISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q1-N106 4. 2176 VL CAGATCGTCCTGACACAGAGCCCAGCTATCATGTCAGCAAGCCCCGGCGAGAAAGTCACAATG- ACTTGCTCAGCCAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCCTCTGGAGTGCCTGCTCACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAAT 5. 2176 L1 SSVSY S27-Y31 6. 2176 L1 TCCTCTGTGAGCTAC -1 7. 2176 L3 QQWSSNPFT Q88-T96 8. 2176 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 9. 2176 L2 DTS D49-S51 10. 2176 L2 GACACATCC -1 11. 2176 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQ Q122- GTTLTVSS S240 12. 2176 VH CAGGTGCAGCTGCAGCAGTCCGGAGCAGAGCTGGCTCGACCAGGAGCTAGTGTGAAAATGTCCTGTAAGGCAA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGGCAGGGACTGGAATGGATCGGGTACATTAATCCTAGCCGAGGATACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCCACTCTGACCACA GATAAGAGCTCCTCTACCGCTTATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCAGTGTACTATTG- CGCCAGGTACTATGACGATCACTACTGTCTGGATTATTGG GGCCAGGGGACTACCCTGACAGTGAGCTCC 13. 2176 H1 GYTFTRYT G147- T154 14. 2176 H1 GGCTACACCTTCACACGGTATACC -1 15. 2176 H3 ARYYDDHYCLDY A218- Y229 16. 2176 H3 GCCAGGTACTATGACGATCACTACTGTCTGGATTAT -1 17. 2176 H2 INPSRGYT I172- T179 18. 2176 H2 ATTAATCCTAGCCGAGGATACACA -1 19. 2176 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 20. 2176 CH2 GCACCAGAGGCTGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGCGTGGTCGTGAGCGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CACTGCCTGCCCCAATCGAGAAGACAATTAGCAAAGC AAAG 21. 2176 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 22. 2176 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTCCTGGACTCA- GATGGGAGCTTCTTTCTGTATAGTAAACTGACCGTGGAC AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAATCTCTGAGTCTGTCACCCGGC 23. 2177 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY SLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 24. 2177 Full CAGATCGTCCTGACACAGAGCCCAGCTATCATGTCAGCAAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CCAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCCTCTGGAGTGCCTGCTCACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGCT- CGACCAGGAGCTAGTGTGAAAATGTCCTGTAAGGC AAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGGCAGGGACTGGAATGGA- TCGGGTACATTAATCCTTCCCGAGGATACACAAACTAC AACCAGAAGTTTAAAGACAAGGCCACTCTGACCACAGATAAGAGCTCCTCTACCGCTTATATGCAGCTGAG- TTCACTGACATCTGAGGACAGTGCAGTGTACTATTGCGC CAGGTACTATGACGATCACTACTCCCTGGATTATTGGGGCCAGGGGACTACCCTGACAGTGAGCTCCGCAG- CCGAACCTAAATCTAGTGACAAGACTCATACCTGCCCCC CTTGTCCAGCACCAGAGGCTGCAGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTG- ATGATCTCCCGGACACCTGAAGTCACTTGTGTGGTCGTG AGCGTGTCTCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCCACATATC GCGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGC- AACAAGGCACTGCCTGCCCCAATCGAGAAGACAATTA GCAAAGCAAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAA- AACCAGGTCAGTCTGCTGTGTCTGGTGAAGGGCTTCTA TCCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTG- TCCTGGACTCAGATGGGAGCTTCTTTCTGTATAGTAAAC TGACCGTGGACAAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCAC- AATCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGC AAG 25. 2177 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q1-N106 26. 2177 VL CAGATCGTCCTGACACAGAGCCCAGCTATCATGTCAGCAAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CCAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCCTCTGGAGTGCCTGCTCACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAAT 27. 2177 L1 SSVSY S27-Y31 28. 2177 L1 TCCTCTGTGAGCTAC -1 29. 2177 L3 QQWSSNPFT Q88-T96 30. 2177 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 31. 2177 L2 DTS D49-S51 32. 2177 L2 GACACATCC -1 33. 2177 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ Q122- GTTLTVSS S240 34. 2177 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGCTCGACCAGGAGCTAGTGTGAAAATGTCCTGTAAGGCAA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGGCAGGGACTGGAATGGATCGGGTACATTAATCCTTCCCGAGGATACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCCACTCTGACCACA GATAAGAGCTCCTCTACCGCTTATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCAGTGTACTATTG- CGCCAGGTACTATGACGATCACTACTCCCTGGATTATTGG GGCCAGGGGACTACCCTGACAGTGAGCTCC 35. 2177 H1 GYTFTRYT G147- T154 36. 2177 H1 GGCTACACCTTCACACGGTATACC -1 37. 2177 H3 ARYYDDHYSLDY A218- Y229 38. 2177 H3 GCCAGGTACTATGACGATCACTACTCCCTGGATTAT -1 39. 2177 H2 INPSRGYT I172- T179 40. 2177 H2 ATTAATCCTTCCCGAGGATACACA -1 41. 2177 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 42. 2177 CH2 GCACCAGAGGCTGCAGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGTGTGGTCGTGAGCGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CACTGCCTGCCCCAATCGAGAAGACAATTAGCAAAGC AAAG 43. 2177 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 44. 2177 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTCCTGGACTCA- GATGGGAGCTTCTTTCTGTATAGTAAACTGACCGTGGAC AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAATCTCTGAGTCTGTCACCCGGC 45. 2309 Full DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQTPGKAPKRWIYDTSKLASGVPSRFSGSGSGTDYTFT- ISSLQPEDIATYYCQQWSSNPFTFGQGTKLQITRTVAAPSVFI -1 FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK- HKVYACEVTHQGLSSPVTKSFNRGEC

46. 2309 Full GATATTCAGATGACCCAGAGCCCAAGCTCCCTGAGTGCCTCAGTGGGCGACCGAGTCACCATCACATGCTCCG- CTTCTAGTTCAGTGTCTTACATGAACTGGTATCAGCA -1 GACTCCAGGGAAGGCACCCAAACGGTGGATCTACGATACCTCAAAGCTGGCCAGCGGAGTGCCCTCCAGAT- TCAGCGGCTCCGGGTCTGGAACAGACTATACTTTTACC ATCAGCTCCCTGCAGCCTGAGGATATTGCTACTTACTATTGCCAGCAGTGGTCTAGTAATCCATTCACTTT- TGGCCAGGGGACCAAGCTGCAGATCACAAGGACTGTGGC CGCTCCCAGCGTCTTCATTTTTCCCCCTAGCGACGAGCAGCTGAAATCTGGCACAGCCAGTGTGGTCTGTC- TGCTGAACAATTTCTACCCTCGCGAAGCAAAGGTGCAGTG GAAAGTCGATAACGCCCTGCAGAGTGGCAACAGCCAGGAGAGCGTGACAGAACAGGACTCCAAGGATTCTA- CTTATAGTCTGTCAAGCACCCTGACACTGTCCAAAGCT GACTACGAGAAGCACAAAGTGTATGCATGCGAAGTCACCCATCAGGGACTGTCCTCTCCTGTGACAAAATC- TTTTAACAGAGGCGAATGT 47. 2309 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQTPGKAPKRWIYDTSKLASGVPSRFSGSGSGTDYTFT- ISSLQPEDIATYYCQQWSSNPFTFGQGTKLQIT D1-T106 48. 2309 VL GATATTCAGATGACCCAGAGCCCAAGCTCCCTGAGTGCCTCAGTGGGCGACCGAGTCACCATCACATGCTCCG- CTTCTAGTTCAGTGTCTTACATGAACTGGTATCAGCA -1 GACTCCAGGGAAGGCACCCAAACGGTGGATCTACGATACCTCAAAGCTGGCCAGCGGAGTGCCCTCCAGAT- TCAGCGGCTCCGGGTCTGGAACAGACTATACTTTTACC ATCAGCTCCCTGCAGCCTGAGGATATTGCTACTTACTATTGCCAGCAGTGGTCTAGTAATCCATTCACTTT- TGGCCAGGGGACCAAGCTGCAGATCACA 49. 2309 L1 SSVSY S27-Y31 50. 2309 L1 AGTTCAGTGTCTTAC -1 51. 2309 L3 QQWSSNPFT Q88-T96 52. 2309 L3 CAGCAGTGGTCTAGTAATCCATTCACT -1 53. 2309 L2 DTS D49-S51 54. 2309 L2 GATACCTCA -1 55. 2309 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R107- C213 56. 2309 CL AGGACTGTGGCCGCTCCCAGCGTCTTCATTTTTCCCCCTAGCGACGAGCAGCTGAAATCTGGCACAGCCAGTG- TGGTCTGTCTGCTGAACAATTTCTACCCTCGCGAAGCA -1 AAGGTGCAGTGGAAAGTCGATAACGCCCTGCAGAGTGGCAACAGCCAGGAGAGCGTGACAGAACAGGACTC- CAAGGATTCTACTTATAGTCTGTCAAGCACCCTGACA CTGTCCAAAGCTGACTACGAGAAGCACAAAGTGTATGCATGCGAAGTCACCCATCAGGGACTGTCCTCTCC- TGTGACAAAATCTTTTAACAGAGGCGAATGT 57. 2310 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKRTVAA -1 PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK- ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 58. 2310 Full GATATTCAGCTGACTCAGTCACCCGCTAGCCTGGCAGTGAGTCTGGGCCAGAGGGCCACCATCAGCTGCAAGG- CTTCACAGAGCGTCGACTACGATGGCGACAGCTACC -1 TGAACTGGTATCAGCAGATCCCTGGGCAGCCCCCTAAACTGCTGATCTACGACGCCTCTAATCTGGTGAGT- GGCATCCCCCCACGCTTCTCCGGCTCTGGGAGTGGAACT GATTTTACCCTGAACATTCACCCCGTGGAGAAGGTCGACGCCGCTACATACCATTGCCAGCAGTCCACAGA- GGACCCCTGGACTTTCGGCGGGGGAACCAAGCTGGAAA TCAAACGGACAGTGGCAGCCCCATCCGTCTTCATTTTTCCTCCATCTGACGAGCAGCTGAAATCAGGGACT- GCTAGCGTGGTCTGTCTGCTGAACAATTTTTACCCAAGAG AAGCAAAGGTGCAGTGGAAAGTCGATAACGCCCTGCAGTCCGGAAATTCTCAGGAGAGTGTGACAGAACAG- GATTCAAAGGACAGCACTTATTCCCTGAGCTCCACCCT GACACTGTCCAAAGCTGATTACGAGAAGCACAAAGTGTATGCATGCGAAGTCACCCATCAGGGACTGTCTA- GTCCCGTGACAAAGTCTTTCAATCGAGGCGAATGT 59. 2310 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 60. 2310 VL GATATTCAGCTGACTCAGTCACCCGCTAGCCTGGCAGTGAGTCTGGGCCAGAGGGCCACCATCAGCTGCAAGG- CTTCACAGAGCGTCGACTACGATGGCGACAGCTACC -1 TGAACTGGTATCAGCAGATCCCTGGGCAGCCCCCTAAACTGCTGATCTACGACGCCTCTAATCTGGTGAGT- GGCATCCCCCCACGCTTCTCCGGCTCTGGGAGTGGAACT GATTTTACCCTGAACATTCACCCCGTGGAGAAGGTCGACGCCGCTACATACCATTGCCAGCAGTCCACAGA- GGACCCCTGGACTTTCGGCGGGGGAACCAAGCTGGAAA TCAAA 61. 2310 L1 QSVDYDGDSY 027-Y36 62. 2310 L1 CAGAGCGTCGACTACGATGGCGACAGCTAC -1 63. 2310 L3 QQSTEDPWT Q93- T101 64. 2310 L3 CAGCAGTCCACAGAGGACCCCTGGACT -1 65. 2310 L2 DAS D54-S56 66. 2310 L2 GACGCCTCT -1 67. 2310 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R112- C218 68. 2310 CL CGGACAGTGGCAGCCCCATCCGTCTTCATTTTTCCTCCATCTGACGAGCAGCTGAAATCAGGGACTGCTAGCG- TGGTCTGTCTGCTGAACAATTTTTACCCAAGAGAAGCA -1 AAGGTGCAGTGGAAAGTCGATAACGCCCTGCAGTCCGGAAATTCTCAGGAGAGTGTGACAGAACAGGATTC- AAAGGACAGCACTTATTCCCTGAGCTCCACCCTGACAC TGTCCAAAGCTGATTACGAGAAGCACAAAGTGTATGCATGCGAAGTCACCCATCAGGGACTGTCTAGTCCC- GTGACAAAGTCTTTCAATCGAGGCGAATGT 69. 2183 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT- CVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFY- PSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 70. 2183 Full GATATTCAGCTGACACAGAGTCCTGCATCACTGGCTGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CCAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGTAAGGCTTCTGGCTATGCATTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACCAACTATAATGGAAAGTTCAAAGGCAAGGCCACACTGACTGCTGACGAGTCAAGCTCCACAG- CCTATATGCAGCTGTCTAGTCTGGCAAGCGAGGATTCC GCCGTGTACTTTTGCGCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCTATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAAC CCAAATCCTCTGATAAGACCCACACATGCCCTCCATGTCCAGCTCCTGAGGCTGCAGGAGGACCAAGCGTG- TTCCTGTTTCCCCCTAAACCTAAGGACACACTGATGATCT CTCGGACACCCGAAGTCACTTGTGTGGTCGTGAGCGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTGG- TACGTGGATGGCGTCGAGGTGCATAATGCCAAAACTAA GCCTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTGTCTGTCCTGACCGTGCTGCATCAGGACTGGC- TGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAA GGCACTGCCAGCCCCCATCGAGAAGACAATTTCCAAAGCAAAGGGCCAGCCTCGAGAACCACAGGTCTATG- TGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCAG GTCTCCCTGACATGTCTGGTGAAGGGATTTTATCCTTCTGATATTGCCGTGGAGTGGGAAAGTAATGGCCA- GCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGA TTCTGACGGGAGTTTCGCTCTGGTCAGTAAACTGACTGTGGATAAGTCACGGTGGCAGCAGGGAAACGTCT- TTAGTTGTTCAGTGATGCACGAGGCACTGCACAATCATT ACACCCAGAAAAGCCTGTCCCTGTCTCCCGGCAAG 71. 2183 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 72. 2183 VL GATATTCAGCTGACACAGAGTCCTGCATCACTGGCTGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CCAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAG 73. 2183 L1 QSVDYDGDSY Q27-Y36 74. 2183 L1 CAGTCAGTGGACTATGATGGCGACTCCTAT -1 75. 2183 L3 QQSTEDPWT Q93- T101 76. 2183 L3 CAGCAGTCTACCGAGGACCCCTGGACA -1 77. 2183 L2 DAS D54-S56 78. 2183 L2 GACGCCTCA -1 79. 2183 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 80. 2183 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGTAAGGCTT- CTGGCTATGCATTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACCAACTAT- AATGGAAAGTTCAAAGGCAAGGCCACACTGACTGCT GACGAGTCAAGCTCCACAGCCTATATGCAGCTGTCTAGTCTGGCAAGCGAGGATTCCGCCGTGTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCTATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 81. 2183 H1 GYAFSSYW G152- W159 82. 2183 H1 GGCTATGCATTTTCTAGTTACTGG -1 83. 2183 H3 ARRETTTVGRYYYAMDY A223- Y239 84. 2183 H3 GCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCTATGGACTAC -1 85. 2183 H2 IWPGDGDT I177- T184 86. 2183 H2 ATTTGGCCCGGGGATGGAGACACC -1 87. 2183 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 88. 2183 CH2 GCTCCTGAGGCTGCAGGAGGACCAAGCGTGTTCCTGTTTCCCCCTAAACCTAAGGACACACTGATGATCTCTC- GGACACCCGAAGTCACTTGTGTGGTCGTGAGCGTGAG -1 CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACTAAGC- CTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTG TCTGTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- ACTGCCAGCCCCCATCGAGAAGACAATTTCCAAAGCA AAG 89. 2183 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 90. 2183 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCAGGTCTCCC- TGACATGTCTGGTGAAGGGATTTTATCCTTCTGATAT -1 TGCCGTGGAGTGGGAAAGTAATGGCCAGCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGATTCTG- ACGGGAGTTTCGCTCTGGTCAGTAAACTGACTGTGGAT AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGTTGTTCAGTGATGCACGAGGCACTGCACAATCATTACAC- CCAGAAAAGCCTGTCCCTGTCTCCCGGC 91. 2184 Full QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ -1 GTTLTV555STGGGGSGGGGSGGGGSDIQIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSP- KRWIYDTSKLASGVPAHFRGSGSGTSYSLTISGMEAEDAATYY CQQWSSNPFTFGSGTKLEINRAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVV- VSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIA- VEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK 92. 2184 Full CAGGTCCAGCTGCAGCAGAGCGGAGCAGAGCTGGCTCGACCAGGAGCTAGTGTGAAAATGTCATGCAAGGCAA- GCGGCTACACCTTCACACGGTATACTATGCACTGG -1 GTGAAACAGAGACCCGGACAGGGCCTGGAATGGATCGGGTACATTAACCCTAGCCGAGGATACACCAACTA- CAACCAGAAGTTTAAAGACAAGGCCACCCTGACCACA GATAAGAGCTCCTCTACAGCTTATATGCAGCTGAGTTCACTGACTTCTGAGGACAGTGCCGTGTACTATTG- TGCTCGGTACTATGACGATCATTACTCCCTGGATTATTGG

GGGCAGGGAACTACCCTGACCGTGAGCTCCTCTAGTACAGGAGGAGGAGGCAGTGGAGGAGGAGGGTCAGG- CGGAGGAGGAAGCGACATCCAGATTGTGCTGACAC AGTCTCCAGCTATCATGTCCGCATCTCCCGGCGAGAAAGTCACTATGACCTGCTCCGCCTCAAGCTCCGTG- TCTTACATGAATTGGTATCAGCAGAAATCAGGAACCAGCC CCAAGAGATGGATCTACGACACATCCAAGCTGGCATCTGGAGTGCCTGCACACTTCAGGGGCAGTGGGTCA- GGAACTAGCTATTCCCTGACCATTAGCGGCATGGAGGC CGAAGATGCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAACCCATTCACATTTGGCAGCGGGACTAAGC- TGGAGATCAATAGGGCAGCCGAACCCAAATCAAGCGAC AAGACACATACTTGCCCCCCTTGTCCAGCTCCAGAAGCTGCAGGAGGACCTTCCGTGTTCCTGTTTCCACC- CAAACCAAAGGATACACTGATGATTAGCCGCACCCCTGAG GTCACATGCGTGGTCGTGAGCGTGAGCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGT- CGAAGTGCATAATGCCAAAACCAAGCCTAGGGAGGAA CAGTACAACAGTACATATAGAGTCGTGTCAGTGCTGACCGTCCTGCACCAGGATTGGCTGAACGGCAAGGA- GTACAAATGCAAGGTGTCCAACAAGGCACTGCCTGCCC CAATCGAGAAGACCATTTCTAAAGCTAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCC- CGGGACGAGCTGACTAAAAACCAGGTCTCTCTGACCTGT CTGGTGAAGGGCTTTTACCCATCTGATATTGCAGTCGAGTGGGAAAGTAATGGGCAGCCCGAGAACAATTA- TAAGACAACTCCCCCTGTGCTGGACTCCGATGGGTCTTT CGCACTGGTCAGCAAACTGACAGTGGATAAGTCCAGATGGCAGCAGGGAAACGTCTTTTCTTGTAGTGTGA- TGCATGAAGCCCTGCACAATCATTACACTCAGAAATCAC TGAGCCTGTCCCCCGGCAAG 93. 2184 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ Q1-S119 GTTLTVSS 94. 2184 VH CAGGTCCAGCTGCAGCAGAGCGGAGCAGAGCTGGCTCGACCAGGAGCTAGTGTGAAAATGTCATGCAAGGCAA- GCGGCTACACCTTCACACGGTATACTATGCACTGG -1 GTGAAACAGAGACCCGGACAGGGCCTGGAATGGATCGGGTACATTAACCCTAGCCGAGGATACACCAACTA- CAACCAGAAGTTTAAAGACAAGGCCACCCTGACCACA GATAAGAGCTCCTCTACAGCTTATATGCAGCTGAGTTCACTGACTTCTGAGGACAGTGCCGTGTACTATTG- TGCTCGGTACTATGACGATCATTACTCCCTGGATTATTGG GGGCAGGGAACTACCCTGACCGTGAGCTCC 95. 2184 H1 GYTFTRYT G26-T33 96. 2184 H1 GGCTACACCTTCACACGGTATACT -1 97. 2184 H3 ARYYDDHYSLDY A97-Y108 98. 2184 H3 GCTCGGTACTATGACGATCATTACTCCCTGGATTAT -1 99. 2184 H2 INPSRGYT I51-T58 100. 2184 H2 ATTAACCCTAGCCGAGGATACACC -1 101. 2184 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q140- N245 102. 2184 VL CAGATTGTGCTGACACAGTCTCCAGCTATCATGTCCGCATCTCCCGGCGAGAAAGTCACTATGACCTGCTCCG- CCTCAAGCTCCGTGTCTTACATGAATTGGTATCAGCAG -1 AAATCAGGAACCAGCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCATCTGGAGTGCCTGCACACTT- CAGGGGCAGTGGGTCAGGAACTAGCTATTCCCTGACCA TTAGCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAACCCATTCACATTT- GGCAGCGGGACTAAGCTGGAGATCAAT 103. 2184 L1 SSVSY S166- Y170 104. 2184 L1 AGCTCCGTGTCTTAC -1 105. 2184 L3 QQWSSNPFT Q227- T235 106. 2184 L3 CAGCAGTGGTCTAGTAACCCATTCACA -1 107. 2184 L2 DTS D188- S190 108. 2184 L2 GACACATCC -1 109. 2184 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A264- K373 110. 2184 CH2 GCTCCAGAAGCTGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTGATGATTAGCC- GCACCCCTGAGGTCACATGCGTGGTCGTGAGCGTGAG -1 CCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCATAATGCCAAAACCAAGC- CTAGGGAGGAACAGTACAACAGTACATATAGAGTCGT GTCAGTGCTGACCGTCCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGTCCAACAAGG- CACTGCCTGCCCCAATCGAGAAGACCATTTCTAAAGCT AAG 111. 2184 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G374- G479 112. 2184 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCCCGGGACGAGCTGACTAAAAACCAGGTCTCTC- TGACCTGTCTGGTGAAGGGCTTTTACCCATCTGATAT -1 TGCAGTCGAGTGGGAAAGTAATGGGCAGCCCGAGAACAATTATAAGACAACTCCCCCTGTGCTGGACTCCG- ATGGGTCTTTCGCACTGGTCAGCAAACTGACAGTGGAT AAGTCCAGATGGCAGCAGGGAAACGTCTTTTCTTGTAGTGTGATGCATGAAGCCCTGCACAATCATTACAC- TCAGAAATCACTGAGCCTGTCCCCCGGC 113. 2185 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT- CVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFY- PSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 114. 2185 Full GATATTCAGCTGACCCAGAGTCCTGCATCACTGGCTGTGAGCCTGGGACAGCGAGCAACAATCTCCTGCAAAG- CCAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCTTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGAACC GATTTTACACTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACAGA- GGACCCCTGGACTTTCGGCGGGGGAACCAAACTGGAAA TCAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGTAAGGCTTCTGGCTATGCATTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACAAACTATAATGGAAAGTTCAAAGGCAAGGCCACTCTGACCGCTGACGAGTCAAGCTCCACTG- CTTATATGCAGCTGTCTAGTCTGGCAAGCGAGGATTCC GCCGTCTACTTTTGCGCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAAC CCAAATCCTCTGATAAGACACACACTTGCCCTCCATGTCCAGCACCTGAGGCTGCAGGAGGACCAAGCGTG- TTCCTGTTTCCCCCTAAACCTAAGGACACTCTGATGATCT CTCGGACTCCCGAAGTCACCTGTGTGGTCGTGAGCGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTGG- TACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAA GCCTAGGGAGGAACAGTATAACTCCACATACCGCGTCGTGTCTGTCCTGACTGTGCTGCATCAGGACTGGC- TGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAA GGCACTGCCAGCCCCCATCGAGAAGACCATTTCCAAAGCCAAGGGCCAGCCTCGAGAACCACAGGTCTATG- TGCTGCCACCCAGCCGGGACGAGCTGACAAAAAACCAG GTCTCCCTGCTGTGTCTGGTGAAGGGATTCTACCCTTCTGATATTGCTGTGGAGTGGGAAAGTAATGGCCA- GCCAGAAAACAATTATCTGACTTGGCCTCCAGTGCTGGA TTCTGACGGGAGTTTCTTTCTGTACAGTAAACTGACCGTGGATAAGTCACGGTGGCAGCAGGGAAACGTCT- TTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATT ACACCCAGAAAAGCCTGTCCCTGTCTCCCGGCAAG 115. 2185 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 116. 2185 VL GATATTCAGCTGACCCAGAGTCCTGCATCACTGGCTGTGAGCCTGGGACAGCGAGCAACAATCTCCTGCAAAG- CCAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCTTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGAACC GATTTTACACTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACAGA- GGACCCCTGGACTTTCGGCGGGGGAACCAAACTGGAAA TCAAG 117. 2185 L1 QSVDYDGDSY Q27-Y36 118. 2185 L1 CAGTCAGTGGACTATGATGGCGACTCCTAT -1 119. 2185 L3 QQSTEDPWT Q93- T101 120. 2185 L3 CAGCAGTCTACAGAGGACCCCTGGACT -1 121. 2185 L2 DAS D54-S56 122. 2185 L2 GACGCTTCA -1 123. 2185 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 124. 2185 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGTAAGGCTT- CTGGCTATGCATTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACAAACTAT- AATGGAAAGTTCAAAGGCAAGGCCACTCTGACCGCT GACGAGTCAAGCTCCACTGCTTATATGCAGCTGTCTAGTCTGGCAAGCGAGGATTCCGCCGTCTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCAATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 125. 2185 H1 GYAFSSYW G152- W159 126. 2185 H1 GGCTATGCATTTTCTAGTTACTGG -1 127. 2185 H3 ARRETTTVGRYYYAMDY A223- Y239 128. 2185 H3 GCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTAC -1 129. 2185 H2 IWPGDGDT I177- T184 130. 2185 H2 ATTTGGCCCGGGGATGGAGACACA -1 131. 2185 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 132. 2185 CH2 GCACCTGAGGCTGCAGGAGGACCAAGCGTGTTCCTGTTTCCCCCTAAACCTAAGGACACTCTGATGATCTCTC- GGACTCCCGAAGTCACCTGTGTGGTCGTGAGCGTGAG -1 CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAAGC- CTAGGGAGGAACAGTATAACTCCACATACCGCGTCGTG TCTGTCCTGACTGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- ACTGCCAGCCCCCATCGAGAAGACCATTTCCAAAGCCA AG 133. 2185 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 134. 2185 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGCTGCCACCCAGCCGGGACGAGCTGACAAAAAACCAGGTCTCCC- TGCTGTGTCTGGTGAAGGGATTCTACCCTTCTGATAT -1 TGCTGTGGAGTGGGAAAGTAATGGCCAGCCAGAAAACAATTATCTGACTTGGCCTCCAGTGCTGGATTCTG- ACGGGAGTTTCTTTCTGTACAGTAAACTGACCGTGGATA AGTCACGGTGGCAGCAGGGAAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACACC- CAGAAAAGCCTGTCCCTGTCTCCCGGC 135. 8074 Full QVQLQQPGAELVKPGTSVKLSCKSSGYTFTSYWIHWVKQRPGHGLEWIGEIDPSDNYTYYNQNFKGKATLTID- KSSSTAYMQLNSLTSEDSAVFYCARSGYGNLFVYWGQGTL -1 VTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS- SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY- RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKS- RWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 136. 8074 Full CAGGTCCAGCTGCAGCAGCCTGGGGCAGAACTGGTCAAACCTGGCACTTCCGTGAAACTGTCATGTAAATCCT- CTGGCTATACTTTTACATCCTACTGGATTCACTGGGTG -1 AAGCAGCGACCAGGACATGGACTGGAGTGGATCGGAGAAATTGACCCTAGTGATAACTACACCTACTACAA- CCAGAATTTCAAGGGCAAAGCAACCCTGACAATTGACA AAAGCTCCTCTACTGCCTACATGCAGCTGAATAGTCTGACCTCAGAGGATAGCGCCGTCTTTTATTGCGCT- AGAAGCGGATACGGCAATCTGTTCGTCTATTGGGGCCAG GGAACCCTGGTCACCGTCTCTGCTGCTAGCACTAAGGGGCCTTCCGTGTTTCCACTGGCTCCCTCTAGTAA- ATCCACCTCTGGAGGCACAGCTGCACTGGGATGTCTGGTG AAGGATTACTTCCCTGAACCAGTCACAGTGAGTTGGAACTCAGGGGCTCTGACAAGTGGAGTCCATACTTT- TCCCGCAGTGCTGCAGTCAAGCGGACTGTACTCCCTGTC CTCTGTGGTCACCGTGCCTAGTTCAAGCCTGGGCACCCAGACATATATCTGCAACGTGAATCACAAGCCAT- CAAATACAAAAGTCGACAAGAAAGTGGAGCCCAAGAGC TGTGATAAAACTCATACCTGCCCACCTTGTCCGGCGCCAGAACTGCTGGGAGGACCAAGCGTGTTCCTGTT- TCCACCCAAGCCTAAAGACACCCTGATGATTTCCCGGACT CCTGAGGTCACCTGCGTGGTCGTGGACGTGTCTCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGA-

TGGCGTCGAAGTGCATAATGCCAAGACCAAACCCCGGG AGGAACAGTACAACTCTACCTATAGAGTCGTGAGTGTCCTGACAGTGCTGCACCAGGACTGGCTGAATGGG- AAGGAGTATAAGTGTAAAGTGAGCAACAAAGCCCTGC CCGCCCCAATCGAAAAAACAATCTCTAAAGCAAAAGGACAGCCTCGCGAACCACAGGTCTACGTGCTGCCC- CCTAGCCGCGACGAACTGACTAAAAATCAGGTCTCTCTG CTGTGTCTGGTCAAAGGATTCTACCCTTCCGACATCGCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAA- CAATTACCTGACCTGGCCCCCTGTGCTGGACTCTGATGG GAGTTTCTTTCTGTATTCAAAGCTGACAGTCGATAAAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCT- CCGTCATGCACGAAGCACTGCACAACCATTACACTCAGA AGTCCCTGTCCCTGTCACCTGGC 137. 8074 VH QVQLQQPGAELVKPGTSVKLSCKSSGYTFTSYWIHWVKQRPGHGLEWIGEIDPSDNYTYYNQNFKGKATLTID- KSSSTAYMQLNSLTSEDSAVFYCARSGYGNLFVYWGQGTL Q1-A118 VTVSA 138. 8074 VH CAGGTCCAGCTGCAGCAGCCTGGGGCAGAACTGGTCAAACCTGGCACTTCCGTGAAACTGTCATGTAAATCCT- CTGGCTATACTTTTACATCCTACTGGATTCACTGGGTG -1 AAGCAGCGACCAGGACATGGACTGGAGTGGATCGGAGAAATTGACCCTAGTGATAACTACACCTACTACAA- CCAGAATTTCAAGGGCAAAGCAACCCTGACAATTGACA AAAGCTCCTCTACTGCCTACATGCAGCTGAATAGTCTGACCTCAGAGGATAGCGCCGTCTTTTATTGCGCT- AGAAGCGGATACGGCAATCTGTTCGTCTATTGGGGCCAG GGAACCCTGGTCACCGTCTCTGCT 139. 8074 H1 KSSGYTFTSYW K23-W33 140. 8074 H1 AAATCCTCTGGCTATACTTTTACATCCTACTGG -1 141. 8074 H3 ARSGYGNLFVY A97-Y107 142. 8074 H3 GCTAGAAGCGGATACGGCAATCTGTTCGTCTAT -1 143. 8074 H2 IDPSDNYT I51-T58 144. 8074 H2 ATTGACCCTAGTGATAACTACACC -1 145. 8074 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A119- V216 146. 8074 CH1 GCTAGCACTAAGGGGCCTTCCGTGTTTCCACTGGCTCCCTCTAGTAAATCCACCTCTGGAGGCACAGCTGCAC- TGGGATGTCTGGTGAAGGATTACTTCCCTGAACCAGTC -1 ACAGTGAGTTGGAACTCAGGGGCTCTGACAAGTGGAGTCCATACTTTTCCCGCAGTGCTGCAGTCAAGCGG- ACTGTACTCCCTGTCCTCTGTGGTCACCGTGCCTAGTTC AAGCCTGGGCACCCAGACATATATCTGCAACGTGAATCACAAGCCATCAAATACAAAAGTCGACAAGAAAG- TG 147. 8074 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A232- K341 148. 8074 CH2 GCGCCAGAACTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAGCCTAAAGACACCCTGATGATTTCCC- GGACTCCTGAGGTCACCTGCGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAAGTGCATAATGCCAAGACCAAAC- CCCGGGAGGAACAGTACAACTCTACCTATAGAGTCGTG AGTGTCCTGACAGTGCTGCACCAGGACTGGCTGAATGGGAAGGAGTATAAGTGTAAAGTGAGCAACAAAGC- CCTGCCCGCCCCAATCGAAAAAACAATCTCTAAAGCAA AA 149. 8074 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G342- G447 150. 8074 CH3 GGACAGCCTCGCGAACCACAGGTCTACGTGCTGCCCCCTAGCCGCGACGAACTGACTAAAAATCAGGTCTCTC- TGCTGTGTCTGGTCAAAGGATTCTACCCTTCCGACATC -1 GCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCTGA- TGGGAGTTTCTTTCTGTATTCAAAGCTGACAGTCGATA AAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCTCCGTCATGCACGAAGCACTGCACAACCATTACACT- CAGAAGTCCCTGTCCCTGTCACCTGGC 151. 3344 Full QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMGVGWIRQPPGKALEWLAHIWWDDDKRYNPALKSRLTISK- DTSKNQVVLTMTNMDPVDTAAYYCARMELWSYYFDYWG -1 QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL- YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY- NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLT- VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 152. 3344 Full CAGGTGACACTGAGAGAATCCGGCCCAGCCCTGGTGAAGCCCACTCAGACCCTGACACTGACTTGCACCTTCT- CTGGGTTTTCCCTGTCTACAAGTGGGATGGGAGTGG -1 GATGGATCAGGCAGCCACCTGGAAAAGCCCTGGAGTGGCTGGCTCACATTTGGTGGGACGATGACAAGCGG- TACAACCCAGCACTGAAAAGCAGACTGACAATCAGCA AGGATACTTCCAAAAACCAGGTGGTCCTGACAATGACTAATATGGACCCCGTGGACACAGCCGCTTACTAT- TGCGCCCGCATGGAACTGTGGAGCTACTATTTCGACTAC TGGGGGCAGGGAACACTGGTCACTGTGAGCTCCGCTTCTACTAAGGGACCCAGTGTGTTTCCTCTGGCACC- ATCTAGTAAATCAACCAGCGGAGGAACAGCAGCACTGG GATGTCTGGTGAAGGATTATTTCCCTGAGCCAGTCACCGTGTCCTGGAACTCTGGCGCACTGACCAGCGGG- GTCCATACATTTCCCGCCGTGCTGCAGTCAAGCGGGCTG TACAGCCTGTCCTCTGTGGTCACCGTGCCTAGTTCAAGCCTGGGAACCCAGACATATATCTGCAACGTGAA- TCACAAGCCCTCTAATACAAAAGTCGATAAGAAAGTGGA ACCTAAGAGTTGTGACAAAACTCATACCTGCCCACCATGTCCAGCACCTGAGCTGCTGGGAGGCCCTTCCG- TGTTCCTGTTTCCTCCAAAGCCAAAAGATACACTGATGAT TAGCCGAACTCCTGAAGTGACCTGCGTGGTCGTGGATGTGAGCCACGAGGACCCAGAAGTCAAGTTCAACT- GGTACGTGGACGGCGTCGAGGTGCATAATGCTAAGAC CAAACCTAGGGAGGAACAGTACAATTCAACATATCGCGTCGTGAGCGTCCTGACTGTGCTGCACCAGGATT- GGCTGAACGGGAAGGAGTATAAGTGCAAAGTGTCCAAT AAGGCTCTGCCAGCACCCATCGAGAAAACCATTTCTAAGGCTAAAGGCCAGCCCCGAGAACCTCAGGTCTA- CGTGTATCCCCCTAGCCGGGATGAGCTGACTAAGAACC AGGTCTCCCTGACCTGTCTGGTGAAAGGATTTTACCCAAGTGACATTGCAGTGGAGTGGGAATCAAATGGC- CAGCCCGAAAACAATTATAAGACCACACCACCCGTGCT GGATAGTGACGGCTCATTCGCCCTGGTCTCCAAGCTGACTGTGGACAAATCTAGGTGGCAGCAGGGGAACG- TCTTTTCCTGTTCTGTGATGCATGAGGCTCTGCACAATC ATTACACCCAGAAGAGTCTGTCACTGAGCCCCGGCAAA 153. 3344 VH QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMGVGWIRQPPGKALEWLAHIWWDDDKRYNPALKSRLTISK- DTSKNQVVLTMTNMDPVDTAAYYCARMELWSYYFDYWG Q1-S120 QGTLVTVSS 154. 3344 VH CAGGTGACACTGAGAGAATCCGGCCCAGCCCTGGTGAAGCCCACTCAGACCCTGACACTGACTTGCACCTTCT- CTGGGTTTTCCCTGTCTACAAGTGGGATGGGAGTGG -1 GATGGATCAGGCAGCCACCTGGAAAAGCCCTGGAGTGGCTGGCTCACATTTGGTGGGACGATGACAAGCGG- TACAACCCAGCACTGAAAAGCAGACTGACAATCAGCA AGGATACTTCCAAAAACCAGGTGGTCCTGACAATGACTAATATGGACCCCGTGGACACAGCCGCTTACTAT- TGCGCCCGCATGGAACTGTGGAGCTACTATTTCGACTAC TGGGGGCAGGGAACACTGGTCACTGTGAGCTCC 155. 3344 H1 GFSLSTSGMG G26-635 156. 3344 H1 GGGTTTTCCCTGTCTACAAGTGGGATGGGA -1 157. 3344 H3 ARMELWSYYFDY A98-Y109 158. 3344 H3 GCCCGCATGGAACTGTGGAGCTACTATTTCGACTAC -1 159. 3344 H2 IWWDDDK I53-K59 160. 3344 H2 ATTTGGTGGGACGATGACAAG -1 161. 3344 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A121- V218 162. 3344 CH1 GCTTCTACTAAGGGACCCAGTGTGTTTCCTCTGGCACCATCTAGTAAATCAACCAGCGGAGGAACAGCAGCAC- TGGGATGTCTGGTGAAGGATTATTTCCCTGAGCCAGT -1 CACCGTGTCCTGGAACTCTGGCGCACTGACCAGCGGGGTCCATACATTTCCCGCCGTGCTGCAGTCAAGCG- GGCTGTACAGCCTGTCCTCTGTGGTCACCGTGCCTAGTT CAAGCCTGGGAACCCAGACATATATCTGCAACGTGAATCACAAGCCCTCTAATACAAAAGTCGATAAGAAA- GTG 163. 3344 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A234- K343 164. 3344 CH2 GCACCTGAGCTGCTGGGAGGCCCTTCCGTGTTCCTGTTTCCTCCAAAGCCAAAAGATACACTGATGATTAGCC- GAACTCCTGAAGTGACCTGCGTGGTCGTGGATGTGAG -1 CCACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCTAAGACCAAAC- CTAGGGAGGAACAGTACAATTCAACATATCGCGTCGT GAGCGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGGAAGGAGTATAAGTGCAAAGTGTCCAATAAGG- CTCTGCCAGCACCCATCGAGAAAACCATTTCTAAGGCT AAA 165. 3344 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G344- G449 166. 3344 CH3 GGCCAGCCCCGAGAACCTCAGGTCTACGTGTATCCCCCTAGCCGGGATGAGCTGACTAAGAACCAGGTCTCCC- TGACCTGTCTGGTGAAAGGATTTTACCCAAGTGACAT -1 TGCAGTGGAGTGGGAATCAAATGGCCAGCCCGAAAACAATTATAAGACCACACCACCCGTGCTGGATAGTG- ACGGCTCATTCGCCCTGGTCTCCAAGCTGACTGTGGAC AAATCTAGGTGGCAGCAGGGGAACGTCTTTTCCTGTTCTGTGATGCATGAGGCTCTGCACAATCATTACAC- CCAGAAGAGTCTGTCACTGAGCCCCGGC 167. 3345 Full QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMGVGWIRQPPGKALEWLAHIWWDDDKRYNPALKSRLTISK- DTSKNQVVLTMTNMDPVDTAAYYCARMELWSYYFDYWG -1 QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL- YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY- NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLT- VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 168. 3345 Full CAGGTGACACTGAGAGAATCCGGACCAGCACTGGTGAAGCCAACCCAGACACTGACTCTGACCTGCACATTCT- CTGGGTTTTCCCTGTCTACAAGTGGGATGGGAGTGG -1 GATGGATCAGGCAGCCACCTGGAAAGGCACTGGAGTGGCTGGCTCACATTTGGTGGGACGATGACAAGCGG- TACAACCCAGCCCTGAAAAGCAGACTGACTATCAGCA AGGATACCTCCAAAAACCAGGTGGTCCTGACTATGACCAATATGGACCCCGTGGACACTGCCGCTTACTAT- TGCGCTCGCATGGAACTGTGGAGCTACTATTTCGACTACT GGGGGCAGGGAACTCTGGTCACCGTGAGCTCCGCTTCTACCAAGGGACCCAGTGTGTTTCCTCTGGCACCA- TCTAGTAAATCAACAAGCGGAGGAACTGCAGCACTGGG ATGTCTGGTGAAGGATTATTTCCCTGAGCCAGTCACCGTGTCCTGGAACTCTGGCGCACTGACAAGCGGGG- TCCATACTTTTCCCGCCGTGCTGCAGTCAAGCGGGCTGT ACAGCCTGTCCTCTGTGGTCACAGTGCCTAGTTCAAGCCTGGGAACACAGACTTATATCTGCAACGTGAAT- CACAAGCCCTCTAATACTAAAGTCGATAAGAAAGTGGAA CCTAAGAGTTGTGACAAAACCCATACATGCCCACCATGTCCAGCTCCTGAGCTGCTGGGAGGACCTTCCGT- GTTCCTGTTTCCTCCAAAGCCAAAAGATACACTGATGATT AGCCGAACCCCTGAAGTCACATGCGTGGTCGTGGATGTGAGCCACGAGGACCCAGAAGTCAAGTTCAACTG- GTACGTGGACGGCGTCGAGGTGCATAATGCCAAGACC AAACCTAGGGAGGAACAGTACAATTCAACCTATCGCGTCGTGAGCGTCCTGACAGTGCTGCACCAGGATTG- GCTGAACGGGAAGGAGTATAAGTGCAAAGTGTCCAAT AAGGCTCTGCCAGCACCCATCGAGAAAACCATTTCTAAGGCAAAAGGCCAGCCCCGAGAACCTCAGGTCTA- CGTGCTGCCCCCTAGCCGGGATGAGCTGACAAAGAACC AGGTCTCCCTGCTGTGTCTGGTGAAAGGATTCTATCCAAGTGACATTGCCGTGGAGTGGGAATCAAATGGC- CAGCCCGAAAACAATTACCTGACTTGGCCACCCGTGCTG GATAGTGACGGCTCATTCTTTCTGTATTCCAAGCTGACCGTGGACAAATCTAGGTGGCAGCAGGGGAACGT- CTTTTCCTGTTCTGTGATGCATGAGGCCCTGCACAATCAT TACACCCAGAAGAGTCTGTCACTGAGCCCCGGCAAA 169. 3345 VH QVTLRESGPALVKPTQTLTLTCTFSGFSLSTSGMGVGWIRQPPGKALEWLAHIWWDDDKRYNPALKSRLTISK- DTSKNQVVLTMTNMDPVDTAAYYCARMELWSYYFDYWG Q1-S120 QGTLVTVSS 170. 3345 VH CAGGTGACACTGAGAGAATCCGGACCAGCACTGGTGAAGCCAACCCAGACACTGACTCTGACCTGCACATTCT- CTGGGTTTTCCCTGTCTACAAGTGGGATGGGAGTGG -1 GATGGATCAGGCAGCCACCTGGAAAGGCACTGGAGTGGCTGGCTCACATTTGGTGGGACGATGACAAGCGG- TACAACCCAGCCCTGAAAAGCAGACTGACTATCAGCA AGGATACCTCCAAAAACCAGGTGGTCCTGACTATGACCAATATGGACCCCGTGGACACTGCCGCTTACTAT- TGCGCTCGCATGGAACTGTGGAGCTACTATTTCGACTACT GGGGGCAGGGAACTCTGGTCACCGTGAGCTCC 171. 3345 H1 GFSLSTSGMG G26-635 172. 3345 H1 GGGTTTTCCCTGTCTACAAGTGGGATGGGA -1 173. 3345 H3 ARMELWSYYFDY A98-Y109 174. 3345 H3 GCTCGCATGGAACTGTGGAGCTACTATTTCGACTAC -1 175. 3345 H2 IWWDDDK I53-K59 176. 3345 H2 ATTTGGTGGGACGATGACAAG -1 177. 3345 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A121- V218 178. 3345 CH1 GCTTCTACCAAGGGACCCAGTGTGTTTCCTCTGGCACCATCTAGTAAATCAACAAGCGGAGGAACTGCAGCAC- TGGGATGTCTGGTGAAGGATTATTTCCCTGAGCCAGT -1

CACCGTGTCCTGGAACTCTGGCGCACTGACAAGCGGGGTCCATACTTTTCCCGCCGTGCTGCAGTCAAGCG- GGCTGTACAGCCTGTCCTCTGTGGTCACAGTGCCTAGTT CAAGCCTGGGAACACAGACTTATATCTGCAACGTGAATCACAAGCCCTCTAATACTAAAGTCGATAAGAAA- GTG 179. 3345 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A234- K343 180. 3345 CH2 GCTCCTGAGCTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCTCCAAAGCCAAAAGATACACTGATGATTAGCC- GAACCCCTGAAGTCACATGCGTGGTCGTGGATGTGAG -1 CCACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAGACCAAAC- CTAGGGAGGAACAGTACAATTCAACCTATCGCGTCGT GAGCGTCCTGACAGTGCTGCACCAGGATTGGCTGAACGGGAAGGAGTATAAGTGCAAAGTGTCCAATAAGG- CTCTGCCAGCACCCATCGAGAAAACCATTTCTAAGGCA AAA 181. 3345 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G344- G449 182. 3345 CH3 GGCCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCCCCTAGCCGGGATGAGCTGACAAAGAACCAGGTCTCCC- TGCTGTGTCTGGTGAAAGGATTCTATCCAAGTGACA -1 TTGCCGTGGAGTGGGAATCAAATGGCCAGCCCGAAAACAATTACCTGACTTGGCCACCCGTGCTGGATAGT- GACGGCTCATTCTTTCTGTATTCCAAGCTGACCGTGGAC AAATCTAGGTGGCAGCAGGGGAACGTCTTTTCCTGTTCTGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAGAGTCTGTCACTGAGCCCCGGC 183. 3346 Full EIVLTQSPATLSLSPGERATLSCSASSSVSYMHWYQQKPGQAPRLLIYDTSKLASGIPARFSGSGSGTDFTLT- ISSLEPEDFAVYYCFQGSVYPFTFGQGTKLEIKRTVAA -1 PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSK- ADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 184. 3346 Full GAAATCGTCCTGACACAGAGTCCTGCTACTCTGTCACTGAGCCCAGGCGAGCGAGCTACCCTGTCCTGCTCTG- CAAGCTCCTCTGTGTCCTACATGCACTGGTATCAGCAG -1 AAGCCAGGACAGGCACCACGACTGCTGATCTACGACACCTCTAAACTGGCAAGTGGCATCCCCGCCAGATT- CAGTGGCTCAGGGAGCGGAACAGACTTTACCCTGACAA TCAGTTCACTGGAGCCCGAAGATTTCGCCGTGTACTATTGCTTTCAGGGCAGCGTCTATCCTTTCACCTTT- GGCCAGGGGACAAAGCTGGAGATCAAAAGGACTGTGGCC GCTCCATCCGTCTTCATTTTTCCCCCTTCTGACGAACAGCTGAAGTCAGGCACAGCTAGCGTGGTCTGTCT- GCTGAACAATTTCTACCCCCGCGAGGCCAAGGTGCAGTGG AAAGTCGATAACGCTCTGCAGTCCGGCAACAGCCAGGAGAGTGTGACTGAACAGGACTCAAAGGATAGCAC- CTATTCCCTGAGCTCCACTCTGACCCTGTCTAAAGCAG ATTACGAGAAGCACAAAGTGTATGCCTGCGAAGTCACACATCAGGGACTGTCTAGTCCCGTGACTAAATCC- TTTAACAGAGGCGAATGT 185. 3346 VL EIVLTQSPATLSLSPGERATLSCSASSSVSYMHWYQQKPGQAPRLLIYDTSKLASGIPARFSGSGSGTDFTLT- ISSLEPEDFAVYYCFQGSVYPFTFGQGTKLEIK E1-K106 186. 3346 VL GAAATCGTCCTGACACAGAGTCCTGCTACTCTGTCACTGAGCCCAGGCGAGCGAGCTACCCTGTCCTGCTCTG- CAAGCTCCTCTGTGTCCTACATGCACTGGTATCAGCAG -1 AAGCCAGGACAGGCACCACGACTGCTGATCTACGACACCTCTAAACTGGCAAGTGGCATCCCCGCCAGATT- CAGTGGCTCAGGGAGCGGAACAGACTTTACCCTGACAA TCAGTTCACTGGAGCCCGAAGATTTCGCCGTGTACTATTGCTTTCAGGGCAGCGTCTATCCTTTCACCTTT- GGCCAGGGGACAAAGCTGGAGATCAAA 187. 3346 L1 SSVSY S27-Y31 188. 3346 L1 TCCTCTGTGTCCTAC -1 189. 3346 L3 FQGSVYPFT F88-T96 190. 3346 L3 TTTCAGGGCAGCGTCTATCCTTTCACC -1 191. 3346 L2 DTS D49-S51 192. 3346 L2 GACACCTCT -1 193. 3346 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R107- C213 194. 3346 CL AGGACTGTGGCCGCTCCATCCGTCTTCATTTTTCCCCCTTCTGACGAACAGCTGAAGTCAGGCACAGCTAGCG- TGGTCTGTCTGCTGAACAATTTCTACCCCCGCGAGGCC -1 AAGGTGCAGTGGAAAGTCGATAACGCTCTGCAGTCCGGCAACAGCCAGGAGAGTGTGACTGAACAGGACTC- AAAGGATAGCACCTATTCCCTGAGCTCCACTCTGACCC TGTCTAAAGCAGATTACGAGAAGCACAAAGTGTATGCCTGCGAAGTCACACATCAGGGACTGTCTAGTCCC- GTGACTAAATCCTTTAACAGAGGCGAATGT 195. 1015 Full EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAD- TSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWG -1 QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS5GL- YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKS CDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTK- PREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSD- GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSL SLSPGK 196. 1015 Full GAGGTGCAGCTGGTGGAAAGCGGAGGAGGACTGGTGCAGCCAGGAGGATCTCTGCGACTGAGTTGCGCCGCTT- CAGGATTCAACATCAAGGACACCTACATTCACTGG -1 GTGCGACAGGCTCCAGGAAAAGGACTGGAGTGGGTGGCTCGAATCTATCCCACTAATGGATACACCCGGTA- TGCCGACTCCGTGAAGGGGAGGTTTACTATTAGCGCC GATACATCCAAAAACACTGCTTACCTGCAGATGAACAGCCTGCGAGCCGAAGATACCGCTGTGTACTATTG- CAGTCGATGGGGAGGAGACGGATTCTACGCTATGGATT ATTGGGGACAGGGGACCCTGGTGACAGTGAGCTCCGCCTCTACCAAGGGCCCCAGTGTGTTTCCCCTGGCT- CCTTCTAGTAAATCCACCTCTGGAGGGACAGCCGCTCTG GGATGTCTGGTGAAGGACTATTTCCCCGAGCCTGTGACCGTGAGTTGGAACTCAGGCGCCCTGACAAGCGG- AGTGCACACTTTTCCTGCTGTGCTGCAGTCAAGCGGGC TGTACTCCCTGTCCTCTGTGGTGACAGTGCCAAGTTCAAGCCTGGGCACACAGACTTATATCTGCAACGTG- AATCATAAGCCCTCAAATACAAAAGTGGACAAGAAAGTG GAGCCCAAGAGCTGTGATAAGACCCACACCTGCCCTCCCTGTCCAGCTCCAGAACTGCTGGGAGGACCTAG- CGTGTTCCTGTTTCCCCCTAAGCCAAAAGACACTCTGAT GATTTCCAGGACTCCCGAGGTGACCTGCGTGGTGGTGGACGTGTCTCACGAGGACCCCGAAGTGAAGTTCA- ACTGGTACGTGGATGGCGTGGAAGTGCATAATGCTAA GACAAAACCAAGAGAGGAACAGTACAACTCCACTTATCGCGTCGTGAGCGTGCTGACCGTGCTGCACCAGG- ACTGGCTGAACGGGAAGGAGTATAAGTGCAAAGTCAG TAATAAGGCCCTGCCTGCTCCAATCGAAAAAACCATCTCTAAGGCCAAAGGCCAGCCAAGGGAGCCCCAGG- TGTACGTGCTGCCACCCAGCAGAGACGAACTGACCAAG AACCAGGTGTCCCTGCTGTGTCTGGTGAAAGGCTTCTATCCTAGTGATATTGCTGTGGAGTGGGAATCAAA- TGGACAGCCAGAGAACAATTACCTGACCTGGCCTCCAGT GCTGGACAGCGATGGCAGCTTCTTCCTGTATTCCAAGCTGACAGTGGATAAATCTCGATGGCAGCAGGGGA- ACGTGTTTAGTTGTTCAGTGATGCATGAAGCCCTGCAC AATCATTACACTCAGAAGAGCCTGTCCCTGTCTCCCGGCAAA 197. 1015 VH EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISAD- TSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWG E1-S120 QGTLVTVSS 198. 1015 VH GAGGTGCAGCTGGTGGAAAGCGGAGGAGGACTGGTGCAGCCAGGAGGATCTCTGCGACTGAGTTGCGCCGCTT- CAGGATTCAACATCAAGGACACCTACATTCACTGG -1 GTGCGACAGGCTCCAGGAAAAGGACTGGAGTGGGTGGCTCGAATCTATCCCACTAATGGATACACCCGGTA- TGCCGACTCCGTGAAGGGGAGGTTTACTATTAGCGCC GATACATCCAAAAACACTGCTTACCTGCAGATGAACAGCCTGCGAGCCGAAGATACCGCTGTGTACTATTG- CAGTCGATGGGGAGGAGACGGATTCTACGCTATGGATT ATTGGGGACAGGGGACCCTGGTGACAGTGAGCTCC 199. 1015 H1 GFNIKDTY G26-Y33 200. 1015 H1 GGATTCAACATCAAGGACACCTAC -1 201. 1015 H3 SRWGGDGFYAMDY S97-Y109 202. 1015 H3 AGTCGATGGGGAGGAGACGGATTCTACGCTATGGATTAT -1 203. 1015 H2 IYPTNGYT I51-T58 204. 1015 H2 ATCTATCCCACTAATGGATACACC -1 205. 1015 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS5GLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A121- V218 206. 1015 CH1 GCCTCTACCAAGGGCCCCAGTGTGTTTCCCCTGGCTCCTTCTAGTAAATCCACCTCTGGAGGGACAGCCGCTC- TGGGATGTCTGGTGAAGGACTATTTCCCCGAGCCTGTG -1 ACCGTGAGTTGGAACTCAGGCGCCCTGACAAGCGGAGTGCACACTTTTCCTGCTGTGCTGCAGTCAAGCGG- GCTGTACTCCCTGTCCTCTGTGGTGACAGTGCCAAGTTC AAGCCTGGGCACACAGACTTATATCTGCAACGTGAATCATAAGCCCTCAAATACAAAAGTGGACAAGAAAG- TG 207. 1015 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A234- K343 208. 1015 CH2 GCTCCAGAACTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCCCCTAAGCCAAAAGACACTCTGATGATTTCCA- GGACTCCCGAGGTGACCTGCGTGGTGGTGGACGTGTC -1 TCACGAGGACCCCGAAGTGAAGTTCAACTGGTACGTGGATGGCGTGGAAGTGCATAATGCTAAGACAAAAC- CAAGAGAGGAACAGTACAACTCCACTTATCGCGTCGT GAGCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGGAAGGAGTATAAGTGCAAAGTCAGTAATAAGG- CCCTGCCTGCTCCAATCGAAAAAACCATCTCTAAGGCC AAA 209. 1015 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G344- G449 210. 1015 CH3 GGCCAGCCAAGGGAGCCCCAGGTGTACGTGCTGCCACCCAGCAGAGACGAACTGACCAAGAACCAGGTGTCCC- TGCTGTGTCTGGTGAAAGGCTTCTATCCTAGTGATA -1 TTGCTGTGGAGTGGGAATCAAATGGACAGCCAGAGAACAATTACCTGACCTGGCCTCCAGTGCTGGACAGC- GATGGCAGCTTCTTCCTGTATTCCAAGCTGACAGTGGA TAAATCTCGATGGCAGCAGGGGAACGTGTTTAGTTGTTCAGTGATGCATGAAGCCCTGCACAATCATTACA- CTCAGAAGAGCCTGTCCCTGTCTCCCGGC 211. 9287 Full QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- NSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ -1 GTLVTVSSVEGGSGGSGGSGGSGGVDDIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKR- LIYDTSKLASGVPSRFSGSGSGTDYTLTISSLQPEDAATYYCQQ WSSNPFTFGQGTKLEIKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVS- HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWE- SNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG 212. 9287 Full CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAACTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCCGTGGAGGGAGGCTCCGGAGGCTCTGGAGGCAGCGGCGGCT- CCGGCGGCGTGGACGATATCCAGATGACCCAGAGC CCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCGCCTCCTCTAGCGTGTCCTA- CATGAACTGGTATCAGCAGAAGCCAGGCAAGGCCCCCAA GCGGCTGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGATTCAGCGGCTCCGGCTCTGGCA- CCGACTATACCCTGACAATCTCCTCTCTGCAGCCCGAGG ATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTTTGGCCAGGGCACAAAGCTGGAG- ATCAAGGCCGCCGAGCCCAAGTCTAGCGACAAGACCCA CACATGCCCACCTTGTCCGGCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAA- AGGATACCCTGATGATCAGCCGCACCCCTGAGGTGACAT GCGTGGTGGTGAGCGTGTCCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTG- CACAATGCCAAGACAAAGCCTCGGGAGGAGCAGTAC AATTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAA- GTGCAAGGTGTCCAATAAGGCCCTGCCTGCCCCAATCG AGAAGACCATCTCTAAGGCCAAGGGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGAC- GAGCTGACAAAGAACCAGGTGAGCCTGCTGTGCCTGGT GAAGGGCTTCTATCCAAGCGATATCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGA- CCTGGCCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTC TGTATTCCAAGCTGACAGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCAC- GAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCT CTGAGCCCCGGC 213. 9287 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- NSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ Q1-S119 GTLVTVSS 214. 9287 VH CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAACTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCC 215. 9287 H1 GYTFTRYT G26-T33 216. 9287 H1 GGCTACACCTTCACAAGGTATACC -1

217. 9287 H3 ARYYDDHYSLDY A97-Y108 218. 9287 H3 GCCCGCTACTATGACGATCACTACAGCCTGGATTAT -1 219. 9287 H2 INPSRGYT I51-T58 220. 9287 H2 ATCAACCCTAGCAGGGGCTACACA -1 221. 9287 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDYTLT- ISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIK D138- K243 222. 9287 VL GATATCCAGATGACCCAGAGCCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCGC- CTCCTCTAGCGTGTCCTACATGAACTGGTATCAGCA -1 GAAGCCAGGCAAGGCCCCCAAGCGGCTGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGAT- TCAGCGGCTCCGGCTCTGGCACCGACTATACCCTGACA ATCTCCTCTCTGCAGCCCGAGGATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTT- TGGCCAGGGCACAAAGCTGGAGATCAAG 223. 9287 L1 SSVSY S164- Y168 224. 9287 L1 TCTAGCGTGTCCTAC -1 225. 9287 L3 QQWSSNPFT Q225- T233 226. 9287 L3 CAGCAGTGGAGCTCCAATCCTTTCACC -1 227. 9287 L2 DTS D186- S188 228. 9287 L2 GATACCAGC -1 229. 9287 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 230. 9287 CH2 GCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAAAGGATACCCTGATGATCAGCC- GCACCCCTGAGGTGACATGCGTGGTGGTGAGCGTGT -1 CCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACAAAG- CCTCGGGAGGAGCAGTACAATTCTACCTATAGAGTGG TGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGTCCAATAAG- GCCCTGCCTGCCCCAATCGAGAAGACCATCTCTAAGG CCAAG 231. 9287 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476 232. 9287 CH3 GGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGACGAGCTGACAAAGAACCAGGTGAGCC- TGCTGTGCCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCT- GATGGCAGCTTCTTTCTGTATTCCAAGCTGACAGTGGAT AAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCACGAGGCCCTGCACAATCACTACAC- CCAGAAGTCCCTGTCTCTGAGCCCCGGC 233. 6689 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQK- FKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHY CLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG 234. 6689 Full CAGATCGTCCTGACTCAGAGCCCCGCTATTATGTCCGCTTCCCCTGGAGAAAAGGTCACTATGACTTGTTCCG- CCTCTAGTTCCGTCTCCTACATGAACTGGTATCAGCAG -1 AAATCTGGAACAAGTCCCAAGCGATGGATCTACGACACTTCCAAGCTGGCATCTGGAGTGCCTGCCCACTT- CCGAGGCAGCGGCTCTGGGACAAGTTATTCACTGACTAT TTCTGGCATGGAGGCCGAAGATGCCGCTACATACTATTGCCAGCAGTGGAGCTCCAACCCATTCACCTTTG- GATGTGGCACAAAGCTGGAGATCAATGGCGGAGGAGGC TCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTCCAGCTGCAGCAGAGCGGAGCAGAACTGGCTAG- ACCAGGAGCCAGTGTGAAAATGTCATGCAAGGCCA GCGGCTACACATTCACTCGGTATACCATGCATTGGGTGAAACAGAGACCAGGACAGTGTCTGGAGTGGATC- GGCTACATTAATCCCAGCAGGGGGTACACAAACTACAA CCAGAAGTTTAAAGACAAGGCAACCCTGACCACCGATAAGTCTAGTTCAACAGCTTATATGCAGCTGAGCT- CCCTGACTTCAGAAGACAGCGCTGTGTACTATTGCGCAC GCTACTATGACGATCACTACTGTCTGGATTATTGGGGGCAGGGAACTACCCTGACCGTGTCTAGTGCAGCC- GAGCCTAAATCAAGCGACAAGACCCATACATGCCCCCCT TGTCCGGCGCCAGAAGCTGCAGGCGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAGGATACTCTGAT- GATTAGCCGAACTCCTGAGGTCACCTGCGTGGTCGTGAG CGTGTCCCACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGATGGGGTCGAAGTGCATAATGCCAAAA- CCAAGCCCAGGGAGGAACAGTACAACTCCACTTATCGC GTCGTGTCTGTCCTGACCGTGCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAATGTAAGGTCTCAAA- TAAGGCTCTGCCCGCCCCTATCGAAAAAACTATCTCAAA GGCAAAAGGCCAGCCTCGCGAACCACAGGTCTACGTGCTGCCCCCTAGCCGCGACGAACTGACTAAAAATC- AGGTCTCTCTGCTGTGTCTGGTCAAAGGATTCTACCCTT CCGACATCGCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTG- GACTCTGATGGGAGTTTCTTTCTGTATTCAAAGCTGACA GTCGATAAAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCTCCGTCATGCACGAAGCACTGCACAACCA- TTACACTCAGAAGTCCCTGTCCCTGTCACCTGGC 235. 6689 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEIN Q1-N106 236. 6689 VL CAGATCGTCCTGACTCAGAGCCCCGCTATTATGTCCGCTTCCCCTGGAGAAAAGGTCACTATGACTTGTTCCG- CCTCTAGTTCCGTCTCCTACATGAACTGGTATCAGCAG -1 AAATCTGGAACAAGTCCCAAGCGATGGATCTACGACACTTCCAAGCTGGCATCTGGAGTGCCTGCCCACTT- CCGAGGCAGCGGCTCTGGGACAAGTTATTCACTGACTAT TTCTGGCATGGAGGCCGAAGATGCCGCTACATACTATTGCCAGCAGTGGAGCTCCAACCCATTCACCTTTG- GATGTGGCACAAAGCTGGAGATCAAT 237. 6689 L1 SSVSY S27-Y31 238. 6689 L1 AGTTCCGTCTCCTAC -1 239. 6689 L3 QQWSSNPFT Q88-T96 240. 6689 L3 CAGCAGTGGAGCTCCAACCCATTCACC -1 241. 6689 L2 DTS D49-S51 242. 6689 L2 GACACTTCC -1 243. 6689 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQ Q122- GTTLTVSS S240 244. 6689 VH CAGGTCCAGCTGCAGCAGAGCGGAGCAGAACTGGCTAGACCAGGAGCCAGTGTGAAAATGTCATGCAAGGCCA- GCGGCTACACATTCACTCGGTATACCATGCATTGG -1 GTGAAACAGAGACCAGGACAGTGTCTGGAGTGGATCGGCTACATTAATCCCAGCAGGGGGTACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCAACCCTGACCACC GATAAGTCTAGTTCAACAGCTTATATGCAGCTGAGCTCCCTGACTTCAGAAGACAGCGCTGTGTACTATTG- CGCACGCTACTATGACGATCACTACTGTCTGGATTATTGG GGGCAGGGAACTACCCTGACCGTGTCTAGT 245. 6689 H1 GYTFTRYT G147- T154 246. 6689 H1 GGCTACACATTCACTCGGTATACC -1 247. 6689 H3 ARYYDDHYCLDY A218- Y229 248. 6689 H3 GCACGCTACTATGACGATCACTACTGTCTGGATTAT -1 249. 6689 H2 INPSRGYT I172- T179 250. 6689 H2 ATTAATCCCAGCAGGGGGTACACA -1 251. 6689 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 252. 6689 CH2 GCGCCAGAAGCTGCAGGCGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAGGATACTCTGATGATTAGCC- GAACTCCTGAGGTCACCTGCGTGGTCGTGAGCGTGTC -1 CCACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGATGGGGTCGAAGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACTTATCGCGTCGT GTCTGTCCTGACCGTGCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAATGTAAGGTCTCAAATAAGG- CTCTGCCCGCCCCTATCGAAAAAACTATCTCAAAGGCA AAA 253. 6689 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 254. 6689 CH3 GGCCAGCCTCGCGAACCACAGGTCTACGTGCTGCCCCCTAGCCGCGACGAACTGACTAAAAATCAGGTCTCTC- TGCTGTGTCTGGTCAAAGGATTCTACCCTTCCGACATC -1 GCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCTGA- TGGGAGTTTCTTTCTGTATTCAAAGCTGACAGTCGATA AAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCTCCGTCATGCACGAAGCACTGCACAACCATTACACT- CAGAAGTCCCTGTCCCTGTCACCTGGC 255. 6690 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY SLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG 256. 6690 Full CAGATCGTCCTGACTCAGAGCCCCGCTATTATGTCCGCAAGCCCTGGAGAGAAAGTGACTATGACCTGTTCCG- CATCTAGTTCCGTGTCCTACATGAACTGGTATCAGCA -1 GAAATCTGGAACAAGTCCCAAGCGATGGATCTACGACACTTCCAAGCTGGCATCTGGAGTGCCTGCCCACT- TCCGAGGCAGCGGCTCTGGGACAAGTTATTCACTGACTA TTAGCGGCATGGAGGCCGAAGATGCCGCTACATACTATTGCCAGCAGTGGAGCTCCAACCCATTCACCTTT- GGATGTGGCACAAAGCTGGAGATCAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTCCAGCTGCAGCAGTCCGGAGCAGAACTGGCT- AGACCAGGAGCCAGTGTGAAAATGTCATGCAAGGC CAGCGGCTACACATTCACTCGGTATACCATGCATTGGGTGAAACAGAGACCAGGACAGTGTCTGGAGTGGA- TCGGCTACATTAATCCCAGCAGGGGGTACACAAACTAC AACCAGAAGTTTAAAGACAAGGCAACCCTGACCACCGATAAGTCTAGTTCAACAGCTTATATGCAGCTGAG- CTCCCTGACTTCAGAAGACAGCGCTGTGTACTATTGCGC ACGCTACTATGACGATCACTACTCCCTGGATTATTGGGGGCAGGGAACTACCCTGACCGTGTCTAGTGCAG- CCGAGCCTAAATCAAGCGACAAGACCCATACATGCCCCC CTTGTCCGGCGCCAGAAGCTGCAGGCGGACCAAGTGTGTTCCTGTTTCCACCCAAACCTAAGGATACTCTG- ATGATTTCTCGAACTCCTGAGGTCACCTGCGTGGTCGTG AGCGTGTCCCACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGATGGGGTCGAAGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCAACTTAT CGCGTCGTGTCTGTCCTGACCGTGCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAATGTAAGGTCTC- AAATAAGGCTCTGCCCGCCCCTATCGAAAAAACTATCTC TAAGGCAAAAGGACAGCCTCGCGAACCACAGGTCTACGTGCTGCCCCCTAGCCGCGACGAACTGACTAAAA- ATCAGGTCTCTCTGCTGTGTCTGGTCAAAGGATTCTACC CTTCCGACATCGCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTG- CTGGACTCTGATGGGAGTTTCTTTCTGTATTCAAAGCTG ACAGTCGATAAAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCTCCGTCATGCACGAAGCACTGCACAA- CCATTACACTCAGAAGTCCCTGTCCCTGTCACCTGGC 257. 6690 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEIN Q1-N106 258. 6690 VL CAGATCGTCCTGACTCAGAGCCCCGCTATTATGTCCGCAAGCCCTGGAGAGAAAGTGACTATGACCTGTTCCG- CATCTAGTTCCGTGTCCTACATGAACTGGTATCAGCA -1 GAAATCTGGAACAAGTCCCAAGCGATGGATCTACGACACTTCCAAGCTGGCATCTGGAGTGCCTGCCCACT- TCCGAGGCAGCGGCTCTGGGACAAGTTATTCACTGACTA TTAGCGGCATGGAGGCCGAAGATGCCGCTACATACTATTGCCAGCAGTGGAGCTCCAACCCATTCACCTTT- GGATGTGGCACAAAGCTGGAGATCAAT 259. 6690 L1 SSVSY S27-Y31 260. 6690 L1 AGTTCCGTGTCCTAC -1 261. 6690 L3 QQWSSNPFT Q88-T96 262. 6690 L3 CAGCAGTGGAGCTCCAACCCATTCACC -1 263. 6690 L2 DTS D49-S51 264. 6690 L2 GACACTTCC -1 265. 6690 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQG Q122- TTLTVSS S240 266. 6690 VH

CAGGTCCAGCTGCAGCAGTCCGGAGCAGAACTGGCTAGACCAGGAGCCAGTGTGAAAATGTCATGCAAGGCCA- GCGGCTACACATTCACTCGGTATACCATGCATTGG -1 GTGAAACAGAGACCAGGACAGTGTCTGGAGTGGATCGGCTACATTAATCCCAGCAGGGGGTACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCAACCCTGACCACC GATAAGTCTAGTTCAACAGCTTATATGCAGCTGAGCTCCCTGACTTCAGAAGACAGCGCTGTGTACTATTG- CGCACGCTACTATGACGATCACTACTCCCTGGATTATTGG GGGCAGGGAACTACCCTGACCGTGTCTAGT 267. 6690 H1 GYTFTRYT G147- T154 268. 6690 H1 GGCTACACATTCACTCGGTATACC -1 269. 6690 H3 ARYYDDHYSLDY A218- Y229 270. 6690 H3 GCACGCTACTATGACGATCACTACTCCCTGGATTAT -1 271. 6690 H2 INPSRGYT I172- T179 272. 6690 H2 ATTAATCCCAGCAGGGGGTACACA -1 273. 6690 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 274. 6690 CH2 GCGCCAGAAGCTGCAGGCGGACCAAGTGTGTTCCTGTTTCCACCCAAACCTAAGGATACTCTGATGATTTCTC- GAACTCCTGAGGTCACCTGCGTGGTCGTGAGCGTGTC -1 CCACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGATGGGGTCGAAGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCAACTTATCGCGTCGT GTCTGTCCTGACCGTGCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAATGTAAGGTCTCAAATAAGG- CTCTGCCCGCCCCTATCGAAAAAACTATCTCTAAGGCAA AA 275. 6690 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 276. 6690 CH3 GGACAGCCTCGCGAACCACAGGTCTACGTGCTGCCCCCTAGCCGCGACGAACTGACTAAAAATCAGGTCTCTC- TGCTGTGTCTGGTCAAAGGATTCTACCCTTCCGACATC -1 GCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCTGA- TGGGAGTTTCTTTCTGTATTCAAAGCTGACAGTCGATA AAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCTCCGTCATGCACGAAGCACTGCACAACCATTACACT- CAGAAGTCCCTGTCCCTGTCACCTGGC 277. 6691 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGCGTKLEIKGGGGS -1 GGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQCLEWIGQIWPGDGDTNYN- GKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRET TTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC- VVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYP- SDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG 278. 6691 Full GATATTCAGCTGACACAGAGCCCCGCATCCCTGGCCGTGAGCCTGGGACAGAGAGCAACTATTTCCTGCAAAG- CCTCACAGAGCGTGGACTATGATGGAGACAGCTATC -1 TGAACTGGTACCAGCAGATCCCAGGCCAGCCCCCTAAACTGCTGATCTACGACGCCAGCAATCTGGTGTCC- GGCATCCCACCCAGGTTCAGTGGATCAGGCAGCGGGAC CGATTTTACACTGAACATTCACCCTGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCCACAG- AGGACCCCTGGACTTTCGGATGTGGCACCAAACTGGAAA TCAAGGGCGGGGGAGGCTCAGGAGGAGGAGGGAGCGGAGGAGGAGGCAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAACTGGTCCGACCTGGAAGCTCCGTGA AAATTTCTTGCAAGGCCAGTGGCTATGCTTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGCGACCAGGA- CAGTGTCTGGAGTGGATCGGGCAGATTTGGCCTGGGGA TGGAGACACCAACTATAATGGAAAGTTCAAAGGCAAGGCAACTCTGACCGCCGACGAATCAAGCTCCACAG- CTTATATGCAGCTGTCTAGTCTGGCTAGTGAGGATTCA GCAGTGTACTTTTGCGCCCGGAGAGAAACCACAACTGTGGGCAGATACTATTACGCAATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAG CCCAAATCCTCTGATAAGACACACACTTGCCCTCCATGTCCGGCGCCAGAAGCTGCAGGCGGACCTTCCGT- GTTCCTGTTTCCCCCTAAACCAAAGGACACTCTGATGATC TCTCGCACTCCAGAGGTCACCTGCGTGGTCGTGTCCGTGTCTCACGAGGACCCCGAAGTCAAATTCAACTG- GTATGTGGACGGGGTCGAAGTGCATAATGCCAAAACAA AGCCTAGGGAGGAACAGTATAACTCTACATACCGCGTCGTGAGTGTCCTGACTGTGCTGCATCAGGATTGG- CTGAATGGCAAGGAGTACAAATGTAAGGTCTCAAATAA GGCTCTGCCCGCCCCTATCGAAAAAACTATCTCTAAAGCTAAAGGCCAGCCTCGCGAACCACAGGTCTACG- TGCTGCCCCCTAGCCGCGACGAACTGACTAAAAATCAGG TCTCTCTGCTGTGTCTGGTCAAAGGATTCTACCCTTCCGACATCGCCGTGGAGTGGGAAAGTAACGGCCAG- CCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGAC TCTGATGGGAGTTTCTTTCTGTATTCAAAGCTGACAGTCGATAAAAGCCGGTGGCAGCAGGGCAATGTGTT- CAGCTGCTCCGTCATGCACGAAGCACTGCACAACCATTA CACTCAGAAGTCCCTGTCCCTGTCACCTGGC 279. 6691 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGCGTKLEIK D1-K111 280. 6691 VL GATATTCAGCTGACACAGAGCCCCGCATCCCTGGCCGTGAGCCTGGGACAGAGAGCAACTATTTCCTGCAAAG- CCTCACAGAGCGTGGACTATGATGGAGACAGCTATC -1 TGAACTGGTACCAGCAGATCCCAGGCCAGCCCCCTAAACTGCTGATCTACGACGCCAGCAATCTGGTGTCC- GGCATCCCACCCAGGTTCAGTGGATCAGGCAGCGGGAC CGATTTTACACTGAACATTCACCCTGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCCACAG- AGGACCCCTGGACTTTCGGATGTGGCACCAAACTGGAAA TCAAG 281. 6691 L1 QSVDYDGDSY Q27-Y36 282. 6691 L1 CAGAGCGTGGACTATGATGGAGACAGCTAT -1 283. 6691 L3 QQSTEDPWT Q93- T101 284. 6691 L3 CAGCAGTCCACAGAGGACCCCTGGACT -1 285. 6691 L2 DAS D54-S56 286. 6691 L2 GACGCCAGC -1 287. 6691 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQCLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 288. 6691 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAACTGGTCCGACCTGGAAGCTCCGTGAAAATTTCTTGCAAGGCCA- GTGGCTATGCTTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGCGACCAGGACAGTGTCTGGAGTGGATCGGGCAGATTTGGCCTGGGGATGGAGACACCAACTAT- AATGGAAAGTTCAAAGGCAAGGCAACTCTGACCGCC GACGAATCAAGCTCCACAGCTTATATGCAGCTGTCTAGTCTGGCTAGTGAGGATTCAGCAGTGTACTTTTG- CGCCCGGAGAGAAACCACAACTGTGGGCAGATACTATTA CGCAATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 289. 6691 H1 GYAFSSYW G152- W159 290. 6691 H1 GGCTATGCTTTTTCTAGTTACTGG -1 291. 6691 H3 ARRETTTVGRYYYAMDY A223- Y239 292. 6691 H3 GCCCGGAGAGAAACCACAACTGTGGGCAGATACTATTACGCAATGGACTAC -1 293. 6691 H2 IWPGDGDT I177- T184 294. 6691 H2 ATTTGGCCTGGGGATGGAGACACC -1 295. 6691 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 296. 6691 CH2 GCGCCAGAAGCTGCAGGCGGACCTTCCGTGTTCCTGTTTCCCCCTAAACCAAAGGACACTCTGATGATCTCTC- GCACTCCAGAGGTCACCTGCGTGGTCGTGTCCGTGTCT -1 CACGAGGACCCCGAAGTCAAATTCAACTGGTATGTGGACGGGGTCGAAGTGCATAATGCCAAAACAAAGCC- TAGGGAGGAACAGTATAACTCTACATACCGCGTCGTG AGTGTCCTGACTGTGCTGCATCAGGATTGGCTGAATGGCAAGGAGTACAAATGTAAGGTCTCAAATAAGGC- TCTGCCCGCCCCTATCGAAAAAACTATCTCTAAAGCTAA A 297. 6691 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 298. 6691 CH3 GGCCAGCCTCGCGAACCACAGGTCTACGTGCTGCCCCCTAGCCGCGACGAACTGACTAAAAATCAGGTCTCTC- TGCTGTGTCTGGTCAAAGGATTCTACCCTTCCGACATC -1 GCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCTGA- TGGGAGTTTCTTTCTGTATTCAAAGCTGACAGTCGATA AAAGCCGGTGGCAGCAGGGCAATGTGTTCAGCTGCTCCGTCATGCACGAAGCACTGCACAACCATTACACT- CAGAAGTCCCTGTCCCTGTCACCTGGC 299. 6692 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGCGTKLEIKGGGGS -1 GGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQCLEWIGQIWPGDGDTNYN- GKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRET TTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTC- VVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYP- SDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG 300. 6692 Full GACATCCAGCTGACACAGAGCCCCGCAAGCCTGGCCGTGAGCCTGGGACAGAGAGCCACTATTTCATGCAAAG- CCTCACAGAGCGTGGACTATGATGGAGACAGCTATC -1 TGAACTGGTACCAGCAGATCCCAGGCCAGCCCCCTAAACTGCTGATCTACGACGCCAGCAATCTGGTGTCC- GGCATCCCACCCAGGTTCAGTGGATCAGGCAGCGGGAC CGATTTTACACTGAACATTCACCCTGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCCACAG- AGGACCCCTGGACTTTCGGATGTGGCACCAAACTGGAAA TCAAGGGCGGGGGAGGCTCAGGAGGAGGAGGGAGCGGAGGAGGAGGCAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAACTGGTCCGACCTGGAAGCTCCGTGA AAATTTCTTGCAAGGCCAGTGGCTATGCTTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGCGACCAGGA- CAGTGTCTGGAGTGGATCGGGCAGATTTGGCCTGGGGA TGGAGACACCAACTATAATGGAAAGTTCAAAGGCAAGGCAACTCTGACCGCCGACGAATCAAGCTCCACAG- CTTATATGCAGCTGTCTAGTCTGGCTAGTGAGGATTCA GCAGTGTACTTTTGCGCCCGGAGAGAAACCACAACTGTGGGCAGATACTATTACGCAATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAG CCCAAATCCTCTGATAAGACACACACTTGCCCTCCATGTCCGGCGCCAGAAGCTGCAGGCGGACCTTCCGT- GTTCCTGTTTCCCCCTAAACCAAAGGACACTCTGATGATC TCTCGCACTCCAGAGGTCACCTGCGTGGTCGTGTCCGTGTCTCACGAGGACCCCGAAGTCAAATTCAACTG- GTATGTGGACGGGGTCGAAGTGCATAATGCCAAAACAA AGCCTAGGGAGGAACAGTATAACTCTACATACCGCGTCGTGAGTGTCCTGACTGTGCTGCATCAGGATTGG- CTGAATGGCAAGGAGTACAAATGTAAGGTGAGCAACA AAGCACTGCCCGCCCCTATCGAAAAAACTATTAGCAAAGCAAAAGGACAGCCTCGCGAACCACAGGTCTAC- GTCTACCCCCCATCAAGAGATGAACTGACAAAAAATCA GGTCTCTCTGACATGCCTGGTCAAAGGATTCTACCCTTCCGACATCGCCGTGGAGTGGGAAAGTAACGGCC- AGCCCGAGAACAATTACAAGACCACACCCCCTGTCCTGG ACTCTGATGGGAGTTTCGCTCTGGTGTCAAAGCTGACCGTCGATAAAAGCCGGTGGCAGCAGGGCAATGTG- TTTAGCTGCTCCGTCATGCACGAAGCCCTGCACAATCA CTACACACAGAAGTCCCTGAGCCTGAGCCCTGGC 301. 6692 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGCGTKLEIK D1-K111 302. 6692 VL GACATCCAGCTGACACAGAGCCCCGCAAGCCTGGCCGTGAGCCTGGGACAGAGAGCCACTATTTCATGCAAAG- CCTCACAGAGCGTGGACTATGATGGAGACAGCTATC -1 TGAACTGGTACCAGCAGATCCCAGGCCAGCCCCCTAAACTGCTGATCTACGACGCCAGCAATCTGGTGTCC- GGCATCCCACCCAGGTTCAGTGGATCAGGCAGCGGGAC CGATTTTACACTGAACATTCACCCTGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCCACAG- AGGACCCCTGGACTTTCGGATGTGGCACCAAACTGGAAA TCAAG 303. 6692 L1 QSVDYDGDSY Q27-Y36 304. 6692 L1 CAGAGCGTGGACTATGATGGAGACAGCTAT -1 305. 6692 L3 QQSTEDPWT Q93- T101 306. 6692 L3 CAGCAGTCCACAGAGGACCCCTGGACT -1 307. 6692 L2 DAS D54-S56 308. 6692 L2 GACGCCAGC -1 309. 6692 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQCLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 310. 6692 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAACTGGTCCGACCTGGAAGCTCCGTGAAAATTTCTTGCAAGGCCA- GTGGCTATGCTTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGCGACCAGGACAGTGTCTGGAGTGGATCGGGCAGATTTGGCCTGGGGATGGAGACACCAACTAT- AATGGAAAGTTCAAAGGCAAGGCAACTCTGACCGCC GACGAATCAAGCTCCACAGCTTATATGCAGCTGTCTAGTCTGGCTAGTGAGGATTCAGCAGTGTACTTTTG- CGCCCGGAGAGAAACCACAACTGTGGGCAGATACTATTA CGCAATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC

311. 6692 H1 GYAFSSYW G152- W159 312. 6692 H1 GGCTATGCTTTTTCTAGTTACTGG -1 313. 6692 H3 ARRETTTVGRYYYAMDY A223- Y239 314. 6692 H3 GCCCGGAGAGAAACCACAACTGTGGGCAGATACTATTACGCAATGGACTAC -1 315. 6692 H2 IWPGDGDT I177- T184 316. 6692 H2 ATTTGGCCTGGGGATGGAGACACC -1 317. 6692 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 318. 6692 CH2 GCGCCAGAAGCTGCAGGCGGACCTTCCGTGTTCCTGTTTCCCCCTAAACCAAAGGACACTCTGATGATCTCTC- GCACTCCAGAGGTCACCTGCGTGGTCGTGTCCGTGTCT -1 CACGAGGACCCCGAAGTCAAATTCAACTGGTATGTGGACGGGGTCGAAGTGCATAATGCCAAAACAAAGCC- TAGGGAGGAACAGTATAACTCTACATACCGCGTCGTG AGTGTCCTGACTGTGCTGCATCAGGATTGGCTGAATGGCAAGGAGTACAAATGTAAGGTGAGCAACAAAGC- ACTGCCCGCCCCTATCGAAAAAACTATTAGCAAAGCAA AA 319. 6692 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 320. 6692 CH3 GGACAGCCTCGCGAACCACAGGTCTACGTCTACCCCCCATCAAGAGATGAACTGACAAAAAATCAGGTCTCTC- TGACATGCCTGGTCAAAGGATTCTACCCTTCCGACAT -1 CGCCGTGGAGTGGGAAAGTAACGGCCAGCCCGAGAACAATTACAAGACCACACCCCCTGTCCTGGACTCTG- ATGGGAGTTTCGCTCTGGTGTCAAAGCTGACCGTCGAT AAAAGCCGGTGGCAGCAGGGCAATGTGTTTAGCTGCTCCGTCATGCACGAAGCCCTGCACAATCACTACAC- ACAGAAGTCCCTGAGCCTGAGCCCTGGC 321. 11175 Full DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIKRTVAAP -1 SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA- DYEKHKVYACEVTHQGL5SPVTKSFNRGEC 322. 11175 Full GACATTCAGCTGACCCAGAGCCCTTCCTCCCTGAGCGCCAGCGTGGGAGACCGGGCCACAATCACCTGCAGGG- CCAGCCAATCCGTGGACTACGAGGGCGACTCCTACC -1 TGAACTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACGACGCCTCCAACCTGGTGTCC- GGCATCCCTTCCAGGTTTAGCGGCTCCGGCAGCGGCAC CGATTTCACCCTGACCATCAGCAGCGTGCAGCCCGAGGACGCTGCCACCTACTACTGCCAGCAGAGCACCG- AGGACCCCTGGACCTTCGGCTGTGGCACAAAGCTGGAG ATCAAGAGGACCGTGGCCGCTCCCTCCGTGTTCATCTTCCCCCCCAGCGATGAACAGCTGAAGTCCGGCAC- AGCTTCCGTGGTGTGCCTGCTCAACAACTTCTACCCCAGG GAAGCCAAGGTGCAGTGGAAAGTTGATAACGCCCTGCAGAGCGGCAACTCCCAGGAGTCCGTGACAGAGCA- GGACAGCAAGGACTCCACCTACTCCCTGTCCTCCACCC TGACCCTGTCCAAGGCCGATTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACACACCAGGGCCTGTCC- TCCCCCGTGACCAAGTCCTTCAACAGGGGCGAG 323. 11175 VL DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIK D1-K111 324. 11175 VL GACATTCAGCTGACCCAGAGCCCTTCCTCCCTGAGCGCCAGCGTGGGAGACCGGGCCACAATCACCTGCAGGG- CCAGCCAATCCGTGGACTACGAGGGCGACTCCTACC -1 TGAACTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACGACGCCTCCAACCTGGTGTCC- GGCATCCCTTCCAGGTTTAGCGGCTCCGGCAGCGGCAC CGATTTCACCCTGACCATCAGCAGCGTGCAGCCCGAGGACGCTGCCACCTACTACTGCCAGCAGAGCACCG- AGGACCCCTGGACCTTCGGCTGTGGCACAAAGCTGGAG ATCAAG 325. 11175 L1 QSVDYEGDSY Q27-Y36 326. 11175 L1 CAATCCGTGGACTACGAGGGCGACTCCTAC -1 327. 11175 L3 QQSTEDPWT Q93- T101 328. 11175 L3 CAGCAGAGCACCGAGGACCCCTGGACC -1 329. 11175 L2 DAS D54-S56 330. 11175 L2 GACGCCTCC -1 331. 11175 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R112- C218 332. 11175 CL AGGACCGTGGCCGCTCCCTCCGTGTTCATCTTCCCCCCCAGCGATGAACAGCTGAAGTCCGGCACAGCTTCCG- TGGTGTGCCTGCTCAACAACTTCTACCCCAGGGAAGC -1 CAAGGTGCAGTGGAAAGTTGATAACGCCCTGCAGAGCGGCAACTCCCAGGAGTCCGTGACAGAGCAGGACA- GCAAGGACTCCACCTACTCCCTGTCCTCCACCCTGACC CTGTCCAAGGCCGATTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACACACCAGGGCCTGTCCTCCCC- CGTGACCAAGTCCTTCAACAGGGGCGAGTGC 333. 1064 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT- CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTYPPSRDELTKNQVSLTCLVKGFY- PSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 334. 1064 Full GACATTCAGCTGACACAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGTAAGGCATCTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACTAACTATAATGGAAAGTTCAAAGGCAAGGCTACACTGACTGCAGACGAGTCAAGCTCCACCG- CTTATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCC GCTGTCTACTTTTGCGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAAC CCAAATCCTCTGATAAGACCCACACATGCCCTCCATGTCCAGCACCTGAGCTGCTGGGAGGACCAAGCGTG- TTCCTGTTTCCACCTAAACCTAAGGACACCCTGATGATCT CTCGGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTGG- TACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAA GCCTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTGTCTGTCCTGACCGTGCTGCATCAGGACTGGC- TGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAA GGCCCTGCCAGCTCCCATCGAGAAGACCATTTCCAAAGCTAAGGGCCAGCCTCGAGAACCACAGGTGTATA- CATACCCACCCAGCCGGGACGAGCTGACCAAAAACCAG GTCTCCCTGACATGTCTGGTGAAGGGATTTTATCCTTCTGATATTGCCGTGGAGTGGGAAAGTAATGGCCA- GCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGA TTCTGACGGGAGTTTCGCACTGGTCAGTAAACTGACAGTGGATAAGTCACGGTGGCAGCAGGGAAACGTCT- TTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCAT TACACTCAGAAAAGCCTGTCCCTGTCTCCCGGCAAG 335. 1064 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 336. 1064 VL GACATTCAGCTGACACAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAG 337. 1064 L1 QSVDYDGDSY Q27-Y36 338. 1064 L1 CAGTCAGTGGACTATGATGGCGACTCCTAT -1 339. 1064 L3 QQSTEDPWT Q93- T101 340. 1064 L3 CAGCAGTCTACCGAGGACCCCTGGACA -1 341. 1064 L2 DAS D54-S56 342. 1064 L2 GACGCCTCA -1 343. 1064 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 344. 1064 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGTAAGGCAT- CTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACTAACTAT- AATGGAAAGTTCAAAGGCAAGGCTACACTGACTGCA GACGAGTCAAGCTCCACCGCTTATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCCGCTGTCTACTTTTG- CGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCAATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 345. 1064 H1 GYAFSSYW G152- W159 346. 1064 H1 GGCTATGCCTTTTCTAGTTACTGG -1 347. 1064 H3 ARRETTTVGRYYYAMDY A223- Y239 348. 1064 H3 GCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTAC -1 349. 1064 H2 IWPGDGDT I177- T184 350. 1064 H2 ATTTGGCCCGGGGATGGAGACACT -1 351. 1064 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 352. 1064 CH2 GCACCTGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCTAAACCTAAGGACACCCTGATGATCTCTC- GGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAG -1 CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAAGC- CTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTG TCTGTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- CCTGCCAGCTCCCATCGAGAAGACCATTTCCAAAGCTA AG 353. 1064 CH3 GQPREPQVYTYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 354. 1064 CH3 GGCCAGCCTCGAGAACCACAGGTGTATACATACCCACCCAGCCGGGACGAGCTGACCAAAAACCAGGTCTCCC- TGACATGTCTGGTGAAGGGATTTTATCCTTCTGATAT -1 TGCCGTGGAGTGGGAAAGTAATGGCCAGCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGATTCTG- ACGGGAGTTTCGCACTGGTCAGTAAACTGACAGTGGAT AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACAC- TCAGAAAAGCCTGTCCCTGTCTCCCGGC 355. 1065 Full DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG -1 TTLTVSSVEGGSGGSGGSGGSGGVDDIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRW- IYDTSKVASGVPYRFSGSGSGTSYSLTISSMEAEDAATYYCQ QWSSNPLTFGAGTKLELKAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV- SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLLCLVKGFYPSDIAVEW- ESNGQPENNYMTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK 356. 1065 Full GATATTAAGCTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCCAGTGTGAAAATGTCATGCAAGACCA- GCGGCTACACATTCACTCGGTATACAATGCACTGG -1 GTGAAGCAGAGACCAGGACAGGGACTGGAATGGATCGGATATATTAACCCTTCCCGAGGCTACACCAACTA- TAATCAGAAGTTTAAAGACAAGGCCACTCTGACCACAG ATAAGAGCTCCTCTACCGCTTACATGCAGCTGAGTTCACTGACAAGTGAGGACTCAGCTGTGTACTATTGC- GCAAGGTACTATGACGATCATTACTGTCTGGATTATTGGG GACAGGGCACTACCCTGACTGTCAGCTCCGTGGAAGGAGGGAGCGGAGGCTCCGGAGGATCTGGCGGGAGT- GGAGGCGTGGACGATATCCAGCTGACCCAGTCCCCA GCAATTATGTCCGCCTCTCCCGGCGAGAAAGTCACCATGACATGCCGCGCTTCTAGTTCAGTGAGCTACAT- GAACTGGTATCAGCAGAAATCAGGCACTAGCCCCAAGAG ATGGATCTACGACACCTCCAAGGTCGCATCTGGGGTGCCTTATAGGTTCAGTGGGTCAGGAAGCGGCACCT- CCTACTCTCTGACAATTAGCTCCATGGAGGCAGAAGAT GCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAATCCACTGACTTTTGGGGCCGGAACCAAACTGGAGCT- GAAGGCAGCCGAACCCAAATCAAGCGACAAGACTCACA CCTGCCCCCCTTGTCCAGCACCCGAACTGCTGGGGGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAG-

GATACACTGATGATCAGCCGGACACCTGAGGTCACTTGC GTGGTCGTGGACGTGAGCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCA- TAATGCTAAAACTAAGCCTAGGGAGGAACAGTACAAT AGTACATATAGAGTCGTGTCAGTGCTGACCGTCCTGCATCAGGATTGGCTGAACGGGAAGGAGTACAAATG- CAAGGTGTCCAACAAGGCCCTGCCTGCTCCAATCGAGA AGACAATTTCTAAAGCCAAGGGCCAGCCCCGAGAACCTCAGGTGTATACACTGCCTCCATCCCGGGACGAG- CTGACTAAAAACCAGGTCTCTCTGCTGTGTCTGGTGAAG GGGTTCTACCCATCTGATATTGCTGTGGAGTGGGAAAGTAATGGACAGCCCGAGAACAATTATATGACCTG- GCCCCCTGTCCTGGACTCCGATGGATCTTTCTTTCTGTAC AGCAAACTGACAGTGGACAAGTCCAGATGGCAGCAGGGCAACGTCTTTAGTTGTTCAGTGATGCACGAGGC- CCTGCACAATCATTACACCCAGAAAAGCCTGTCCCTGT CTCCCGGCAAG 357. 1065 VH DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG D1-S119 TTLTVSS 358. 1065 VH GATATTAAGCTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCCAGTGTGAAAATGTCATGCAAGACCA- GCGGCTACACATTCACTCGGTATACAATGCACTGG -1 GTGAAGCAGAGACCAGGACAGGGACTGGAATGGATCGGATATATTAACCCTTCCCGAGGCTACACCAACTA- TAATCAGAAGTTTAAAGACAAGGCCACTCTGACCACAG ATAAGAGCTCCTCTACCGCTTACATGCAGCTGAGTTCACTGACAAGTGAGGACTCAGCTGTGTACTATTGC- GCAAGGTACTATGACGATCATTACTGTCTGGATTATTGGG GACAGGGCACTACCCTGACTGTCAGCTCC 359. 1065 H1 GYTFTRYT G26-T33 360. 1065 H1 GGCTACACATTCACTCGGTATACA -1 361. 1065 H3 ARYYDDHYCLDY A97-Y108 362. 1065 H3 GCAAGGTACTATGACGATCATTACTGTCTGGATTAT -1 363. 1065 H2 INPSRGYT I51-T58 364. 1065 H2 ATTAACCCTTCCCGAGGCTACACC -1 365. 1065 VL DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFSGSGSGTSYSLT- ISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK D138- K243 366. 1065 VL GATATCCAGCTGACCCAGTCCCCAGCAATTATGTCCGCCTCTCCCGGCGAGAAAGTCACCATGACATGCCGCG- CTTCTAGTTCAGTGAGCTACATGAACTGGTATCAGCA -1 GAAATCAGGCACTAGCCCCAAGAGATGGATCTACGACACCTCCAAGGTCGCATCTGGGGTGCCTTATAGGT- TCAGTGGGTCAGGAAGCGGCACCTCCTACTCTCTGACA ATTAGCTCCATGGAGGCAGAAGATGCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAATCCACTGACTTT- TGGGGCCGGAACCAAACTGGAGCTGAAG 367. 1065 L1 SSVSY S164- Y168 368. 1065 L1 AGTTCAGTGAGCTAC -1 369. 1065 L3 QQWSSNPLT Q225- T233 370. 1065 L3 CAGCAGTGGTCTAGTAATCCACTGACT -1 371. 1065 L2 DTS D186- S188 372. 1065 L2 GACACCTCC -1 373. 1065 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 374. 1065 CH2 GCACCCGAACTGCTGGGGGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTGATGATCAGCC- GGACACCTGAGGTCACTTGCGTGGTCGTGGACGTGA GCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCATAATGCTAAAACTAAG- CCTAGGGAGGAACAGTACAATAGTACATATAGAGTCG -1 TGTCAGTGCTGACCGTCCTGCATCAGGATTGGCTGAACGGGAAGGAGTACAAATGCAAGGTGTCCAACAAG- GCCCTGCCTGCTCCAATCGAGAAGACAATTTCTAAAGC CAAG 375. 1065 CH3 GQPREPQVYTLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYMTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476 376. 1065 CH3 GGCCAGCCCCGAGAACCTCAGGTGTATACACTGCCTCCATCCCGGGACGAGCTGACTAAAAACCAGGTCTCTC- TGCTGTGTCTGGTGAAGGGGTTCTACCCATCTGATAT -1 TGCTGTGGAGTGGGAAAGTAATGGACAGCCCGAGAACAATTATATGACCTGGCCCCCTGTCCTGGACTCCG- ATGGATCTTTCTTTCTGTACAGCAAACTGACAGTGGACA AGTCCAGATGGCAGCAGGGCAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACACC- CAGAAAAGCCTGTCCCTGTCTCCCGGC 377. 1067 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY CLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYMTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK 378. 1067 Full CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTCCAGCTGCAGCAGTCCGGAGCTGAGCTGGCA- CGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGC CAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGGCAGGGACTGGAATGGA- TCGGGTACATTAATCCTAGCCGAGGATACACAAACTAC AACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAGATAAGAGCTCCTCTACCGCATATATGCAGCTGAG- TTCACTGACATCTGAGGACAGTGCCGTGTACTATTGCGC TAGGTACTATGACGATCACTACTGTCTGGATTATTGGGGCCAGGGGACTACCCTGACCGTGAGCTCCGCAG- CCGAACCTAAATCTAGTGACAAGACTCATACCTGCCCCC CTTGTCCAGCACCAGAGCTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTG- ATGATCTCCCGGACACCTGAAGTCACTTGCGTGGTCGTG GACGTGTCTCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCCACATATC GCGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGC- AACAAGGCCCTGCCTGCTCCAATCGAGAAGACAATTAG CAAAGCCAAGGGGCAGCCCCGAGAACCTCAGGTGTACACTCTGCCTCCATCTCGGGACGAGCTGACCAAAA- ACCAGGTCAGTCTGCTGTGTCTGGTGAAGGGCTTCTAT CCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACATGACATGGCCCCCTGT- CCTGGACTCAGATGGGAGCTTCTTTCTGTATAGTAAACT GACTGTGGACAAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACA- ATCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGCA AG 379. 1067 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q1-N106 380. 1067 VL CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAAT 381. 1067 L1 SSVSY S27-Y31 382. 1067 L1 TCCTCTGTGAGCTAC -1 383. 1067 L3 QQWSSNPFT Q88-T96 384. 1067 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 385. 1067 L2 DTS D49-S51 386. 1067 L2 GACACATCC -1 387. 1067 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQ Q122- GTTLTVSS S240 388. 1067 VH CAGGTCCAGCTGCAGCAGTCCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGCCA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGGCAGGGACTGGAATGGATCGGGTACATTAATCCTAGCCGAGGATACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCTACTCTGACCACA GATAAGAGCTCCTCTACCGCATATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCCGTGTACTATTG- CGCTAGGTACTATGACGATCACTACTGTCTGGATTATTGG GGCCAGGGGACTACCCTGACCGTGAGCTCC 389. 1067 H1 GYTFTRYT G147- T154 390. 1067 H1 GGCTACACCTTCACACGGTATACC -1 391. 1067 H3 ARYYDDHYCLDY A218- Y229 392. 1067 H3 GCTAGGTACTATGACGATCACTACTGTCTGGATTAT -1 393. 1067 H2 INPSRGYT I172- T179 394. 1067 H2 ATTAATCCTAGCCGAGGATACACA -1 395. 1067 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 396. 1067 CH2 GCACCAGAGCTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGCGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CCCTGCCTGCTCCAATCGAGAAGACAATTAGCAAAGCC AAG 397. 1067 CH3 GQPREPQVYTLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYMTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 398. 1067 CH3 GGGCAGCCCCGAGAACCTCAGGTGTACACTCTGCCTCCATCTCGGGACGAGCTGACCAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGCTTCTATCCAAGCGATAT -1 TGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACATGACATGGCCCCCTGTCCTGGACTCAG- ATGGGAGCTTCTTTCTGTATAGTAAACTGACTGTGGACA AGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACACC- CAGAAATCTCTGAGTCTGTCACCCGGC 399. 3357 Full DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTL- SINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVA -1 APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS- KADYEKHKVYACEVTHQGL5SPVTKSFNRGEC 400. 3357 Full GACATCCTGCTGACTCAGAGCCCAGTGATCCTGTCAGTCAGCCCAGGAGAGCGGGTGTCCTTCTCTTGCAGAG- CAAGTCAGTCAATCGGAACAAATATTCACTGGTACCA -1 GCAGAGGACTAACGGCTCCCCTCGCCTGCTGATTAAGTATGCTAGCGAATCCATCTCTGGCATTCCATCTC- GGTTCAGTGGCTCAGGGAGCGGAACAGACTTTACTCTGT CCATCAATTCTGTGGAGAGTGAAGACATTGCCGATTACTATTGCCAGCAGAACAATAACTGGCCCACCACA- TTCGGCGCTGGGACCAAGCTGGAGCTGAAACGAACAGT GGCCGCTCCTTCTGTCTTCATCTTTCCCCCTAGTGACGAACAGCTGAAAAGCGGCACAGCCTCCGTGGTCT- GTCTGCTGAATAACTTTTACCCAAGAGAGGCAAAGGTGCA GTGGAAAGTCGATAATGCCCTGCAGTCAGGGAACAGCCAGGAGTCCGTGACTGAACAGGACTCTAAGGATA- GTACCTATTCACTGAGCTCCACTCTGACCCTGTCCAAA GCTGATTACGAGAAGCACAAAGTGTATGCATGCGAAGTCACCCATCAGGGGCTGTCTAGTCCCGTGACAAA- GAGCTTTAACCGGGGAGAGTGT 401. 3357 VL DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTL- SINSVESEDIADYYCQQNNNWPTTFGAGTKLELK D1-K107 402. 3357 VL GACATCCTGCTGACTCAGAGCCCAGTGATCCTGTCAGTCAGCCCAGGAGAGCGGGTGTCCTTCTCTTGCAGAG- CAAGTCAGTCAATCGGAACAAATATTCACTGGTACCA -1 GCAGAGGACTAACGGCTCCCCTCGCCTGCTGATTAAGTATGCTAGCGAATCCATCTCTGGCATTCCATCTC- GGTTCAGTGGCTCAGGGAGCGGAACAGACTTTACTCTGT CCATCAATTCTGTGGAGAGTGAAGACATTGCCGATTACTATTGCCAGCAGAACAATAACTGGCCCACCACA- TTCGGCGCTGGGACCAAGCTGGAGCTGAAA 403. 3357 L1 QSIGTN Q27-N32 404. 3357 L1 CAGTCAATCGGAACAAAT -1 405. 3357 L3 QQNNNWPTT Q89-T97

406. 3357 L3 CAGCAGAACAATAACTGGCCCACCACA -1 407. 3357 L2 YAS Y50-S52 408. 3357 L2 TATGCTAGC -1 409. 3357 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R108- C214 410. 3357 CL CGAACAGTGGCCGCTCCTTCTGTCTTCATCTTTCCCCCTAGTGACGAACAGCTGAAAAGCGGCACAGCCTCCG- TGGTCTGTCTGCTGAATAACTTTTACCCAAGAGAGGCA -1 AAGGTGCAGTGGAAAGTCGATAATGCCCTGCAGTCAGGGAACAGCCAGGAGTCCGTGACTGAACAGGACTC- TAAGGATAGTACCTATTCACTGAGCTCCACTCTGACCC TGTCCAAAGCTGATTACGAGAAGCACAAAGTGTATGCATGCGAAGTCACCCATCAGGGGCTGTCTAGTCCC- GTGACAAAGAGCTTTAACCGGGGAGAGTGT 411. 1842 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT- CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFY- PSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 412. 1842 Full GATATTCAGCTGACACAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGTAAGGCATCTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACCAACTATAATGGAAAGTTCAAAGGCAAGGCTACACTGACTGCAGACGAGTCAAGCTCCACAG- CTTATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCC GCTGTGTACTTTTGCGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAA CCCAAATCCTCTGATAAGACCCACACATGCCCTCCATGTCCAGCACCTGAGCTGCTGGGAGGACCAAGCGT- GTTCCTGTTTCCACCTAAACCTAAGGACACACTGATGATC TCTCGGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTG- GTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACTA AGCCTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTGTCTGTCCTGACCGTGCTGCATCAGGACTGG- CTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACA AGGCCCTGCCAGCTCCCATCGAGAAGACAATTTCCAAAGCTAAGGGCCAGCCTCGAGAACCACAGGTCTAT- GTGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCA GGTCTCCCTGACATGTCTGGTGAAGGGATTTTATCCTTCTGATATTGCCGTGGAGTGGGAAAGTAATGGCC- AGCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGG ATTCTGACGGGAGTTTCGCACTGGTCAGTAAACTGACTGTGGATAAGTCACGGTGGCAGCAGGGAAACGTC- TTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCA TTACACCCAGAAAAGCCTGTCCCTGTCTCCCGGCAAG 413. 1842 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 414. 1842 VL GATATTCAGCTGACACAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAG 415. 1842 L1 QSVDYDGDSY Q27-Y36 416. 1842 L1 CAGTCAGTGGACTATGATGGCGACTCCTAT -1 417. 1842 L3 QQSTEDPWT Q93- T101 418. 1842 L3 CAGCAGTCTACCGAGGACCCCTGGACA -1 419. 1842 L2 DAS D54-S56 420. 1842 L2 GACGCCTCA -1 421. 1842 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 422. 1842 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGTAAGGCAT- CTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACCAACTAT- AATGGAAAGTTCAAAGGCAAGGCTACACTGACTGCA GACGAGTCAAGCTCCACAGCTTATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCCGCTGTGTACTTTTG- CGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCAATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 423. 1842 H1 GYAFSSYW G152- W159 424. 1842 H1 GGCTATGCCTTTTCTAGTTACTGG -1 425. 1842 H3 ARRETTTVGRYYYAMDY A223- Y239 426. 1842 H3 GCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTAC -1 427. 1842 H2 IWPGDGDT I177- T184 428. 1842 H2 ATTTGGCCCGGGGATGGAGACACC -1 429. 1842 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 430. 1842 CH2 GCACCTGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCTAAACCTAAGGACACACTGATGATCTCTC- GGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAG CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACTAAGC- CTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTG TCTGTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- CCTGCCAGCTCCCATCGAGAAGACAATTTCCAAAGCTA AG -1 431. 1842 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 432. 1842 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCAGGTCTCCC- TGACATGTCTGGTGAAGGGATTTTATCCTTCTGATAT -1 TGCCGTGGAGTGGGAAAGTAATGGCCAGCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGATTCTG- ACGGGAGTTTCGCACTGGTCAGTAAACTGACTGTGGAT AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACAC- CCAGAAAAGCCTGTCCCTGTCTCCCGGC 433. 2227 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY CLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 434. 2227 Full CAGATCGTCCTGACACAGTCCCCAGCAATCATGTCAGCCAGCCCCGGGGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGGACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTAGCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATGTGGCACCAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTGCAGCTGCAGCAGTCCGGAGCTGAGCTGGCA- CGACCAGGAGCAAGTGTGAAAATGTCATGCAAGGC CAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGACAGTGTCTGGAATGGA- TCGGCTACATTAATCCTTCTCGAGGGTACACAAACTACA ACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAGATAAGAGCTCCTCTACCGCATATATGCAGCTGAGT- TCACTGACATCTGAGGACAGTGCCGTGTACTATTGCGCT AGGTACTATGACGATCACTACTGTCTGGATTATTGGGGGCAGGGAACTACCCTGACAGTGAGCTCCGCAGC- CGAACCTAAATCTAGTGACAAGACTCATACCTGCCCCCC TTGTCCAGCACCAGAGCTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGA- TGATCTCCCGGACACCTGAAGTCACTTGCGTGGTCGTGG ACGTGTCTCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAA- ACCAAGCCCAGGGAGGAACAGTACAACTCCACATATCG CGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCA- ACAAGGCCCTGCCTGCTCCAATCGAGAAGACAATTAGC AAAGCCAAGGGCCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAA- CCAGGTCAGTCTGCTGTGTCTGGTGAAGGGATTCTATC CAAGCGATATTGCTGTGGAGTGGGAATCCAATGGCCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTC- CTGGACTCAGATGGCAGCTTCTTTCTGTATAGTAAACTG ACCGTGGACAAGTCACGGTGGCAGCAGGGGAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAA- TCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGCAA G 435. 2227 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEIN Q1-N106 436. 2227 VL CAGATCGTCCTGACACAGTCCCCAGCAATCATGTCAGCCAGCCCCGGGGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGGACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTAGCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATGTGGCACCAAGCTGGAAATTAAT 437. 2227 L1 SSVSY S27-Y31 438. 2227 L1 TCCTCTGTGAGCTAC -1 439. 2227 L3 QQWSSNPFT Q88-T96 440. 2227 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 441. 2227 L2 DTS D49-S51 442. 2227 L2 GACACATCC -1 443. 2227 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQ Q122- GTTLTVSS S240 444. 2227 VH CAGGTGCAGCTGCAGCAGTCCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCATGCAAGGCCA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGACAGTGTCTGGAATGGATCGGCTACATTAATCCTTCTCGAGGGTACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAG ATAAGAGCTCCTCTACCGCATATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCCGTGTACTATTGC- GCTAGGTACTATGACGATCACTACTGTCTGGATTATTGGG GGCAGGGAACTACCCTGACAGTGAGCTCC 445. 2227 H1 GYTFTRYT G147- T154 446. 2227 H1 GGCTACACCTTCACACGGTATACC -1 447. 2227 H3 ARYYDDHYCLDY A218- Y229 448. 2227 H3 GCTAGGTACTATGACGATCACTACTGTCTGGATTAT -1 449. 2227 H2 INPSRGYT I172- T179 450. 2227 H2 ATTAATCCTTCTCGAGGGTACACA -1 451. 2227 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 452. 2227 CH2 GCACCAGAGCTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGCGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CCCTGCCTGCTCCAATCGAGAAGACAATTAGCAAAGC CAAG 453. 2227 CH3

GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 454. 2227 CH3 GGCCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGATTCTATCCAAGCGATAT -1 TGCTGTGGAGTGGGAATCCAATGGCCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTCCTGGACTCAG- ATGGCAGCTTCTTTCTGTATAGTAAACTGACCGTGGACA AGTCACGGTGGCAGCAGGGGAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACACC- CAGAAATCTCTGAGTCTGTCACCCGGC 455. 2228 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY SLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 456. 2228 Full CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGGGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGC AGAAAAGCGGGACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACAC- TTCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGAC AATTTCCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTT- TTGGATGTGGCACCAAGCTGGAAATTAATGGCGGAGGA GGCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTGCAGCTGCAGCAGTCCGGAGCTGAGCTGGC- ACGACCAGGAGCAAGTGTGAAAATGTCATGCAAGG CCAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGACAGTGTCTGGAATGG- ATCGGCTACATTAATCCTAGCCGAGGGTACACAAACTA CAACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAGATAAGAGCTCCTCTACCGCATATATGCAGCTGA- GTTCACTGACATCTGAGGACAGTGCCGTGTACTATTGCG CTAGGTACTATGACGATCACTACTCCCTGGATTATTGGGGGCAGGGAACTACCCTGACAGTGAGCTCCGCA- GCCGAACCTAAATCTAGTGACAAGACTCATACCTGCCCA CCTTGTCCAGCACCAGAGCTGCTGGGCGGGCCTTCTGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCT- GATGATCTCCCGGACACCTGAAGTCACTTGTGTGGTCGTG GACGTGTCTCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCCACATATC GCGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGC- AACAAGGCCCTGCCTGCTCCAATCGAGAAGACAATTAG CAAAGCCAAGGGCCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAA- ACCAGGTCAGTCTGCTGTGTCTGGTGAAGGGATTCTAT CCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGCCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGT- CCTGGACTCAGATGGCAGCTTCTTTCTGTATAGTAAACT GACCGTGGACAAGTCACGGTGGCAGCAGGGGAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACA- ATCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGC AAG -1 457. 2228 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGCGTKLEIN Q1-N106 458. 2228 VL CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGGGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGC -1 AGAAAAGCGGGACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACAC- TTCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGAC AATTTCCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTT- TTGGATGTGGCACCAAGCTGGAAATTAAT 459. 2228 L1 SSVSY S27-Y31 460. 2228 L1 TCCTCTGTGAGCTAC -1 461. 2228 L3 QQWSSNPFT Q88-T96 462. 2228 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 463. 2228 L2 DTS D49-S51 464. 2228 L2 GACACATCC -1 465. 2228 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQCLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQG Q122- TTLTVSS S240 466. 2228 VH CAGGTGCAGCTGCAGCAGTCCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCATGCAAGGCCA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGACAGTGTCTGGAATGGATCGGCTACATTAATCCTAGCCGAGGGTACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAG ATAAGAGCTCCTCTACCGCATATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCCGTGTACTATTGC- GCTAGGTACTATGACGATCACTACTCCCTGGATTATTGGG GGCAGGGAACTACCCTGACAGTGAGCTCC 467. 2228 H1 GYTFTRYT G147- T154 468. 2228 H1 GGCTACACCTTCACACGGTATACC -1 469. 2228 H3 ARYYDDHYSLDY A218- Y229 470. 2228 H3 GCTAGGTACTATGACGATCACTACTCCCTGGATTAT -1 471. 2228 H2 INPSRGYT I172- T179 472. 2228 H2 ATTAATCCTAGCCGAGGGTACACA -1 473. 2228 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 474. 2228 CH2 GCACCAGAGCTGCTGGGCGGGCCTTCTGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGTGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CCCTGCCTGCTCCAATCGAGAAGACAATTAGCAAAGC CAAG 475. 2228 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 476. 2228 CH3 GGCCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGATTCTATCCAAGCGATAT -1 TGCTGTGGAGTGGGAATCCAATGGCCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTCCTGGACTCAG- ATGGCAGCTTCTTTCTGTATAGTAAACTGACCGTGGACA AGTCACGGTGGCAGCAGGGGAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACACC- CAGAAATCTCTGAGTCTGTCACCCGGC 477. 1844 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVT- CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFY- PSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 478. 1844 Full GATATTCAGCTGACACAGAGTCCTGCATCACTGGCTGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CCAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGTAAGGCTTCTGGCTATGCATTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACCAACTATAATGGAAAGTTCAAAGGCAAGGCCACACTGACTGCTGACGAGTCAAGCTCCACAG- CCTATATGCAGCTGTCTAGTCTGGCAAGCGAGGATTCC GCCGTGTACTTTTGCGCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCTATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAAC CCAAATCCTCTGATAAGACCCACACATGCCCTCCATGTCCAGCTCCTGAGGCTGCAGGAGGACCAAGCGTG- TTCCTGTTTCCCCCTAAACCTAAGGACACACTGATGATCT CTCGGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTGG- TACGTGGATGGCGTCGAGGTGCATAATGCCAAAACTAA GCCTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTGTCTGTCCTGACCGTGCTGCATCAGGACTGGC- TGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAA GGCACTGCCAGCCCCCATCGAGAAGACAATTTCCAAAGCAAAGGGCCAGCCTCGAGAACCACAGGTCTATG- TGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCAG GTCTCCCTGACATGTCTGGTGAAGGGATTTTATCCTTCTGATATTGCCGTGGAGTGGGAAAGTAATGGCCA- GCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGA TTCTGACGGGAGTTTCGCTCTGGTCAGTAAACTGACTGTGGATAAGTCACGGTGGCAGCAGGGAAACGTCT- TTAGTTGTTCAGTGATGCACGAGGCACTGCACAATCATT ACACCCAGAAAAGCCTGTCCCTGTCTCCCGGCAAG 479. 1844 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 480. 1844 VL GATATTCAGCTGACACAGAGTCCTGCATCACTGGCTGTGAGCCTGGGACAGCGAGCAACTATCTCCTGCAAAG- CCAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGCGGGGGAACTAAACTGGAAA TCAAG 481. 1844 L1 QSVDYDGDSY Q27-Y36 482. 1844 L1 CAGTCAGTGGACTATGATGGCGACTCCTAT -1 483. 1844 L3 QQSTEDPWT Q93- T101 484. 1844 L3 CAGCAGTCTACCGAGGACCCCTGGACA -1 485. 1844 L2 DAS D54-S56 486. 1844 L2 GACGCCTCA -1 487. 1844 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 488. 1844 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGTAAGGCTT- CTGGCTATGCATTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACCAACTAT- AATGGAAAGTTCAAAGGCAAGGCCACACTGACTGCT GACGAGTCAAGCTCCACAGCCTATATGCAGCTGTCTAGTCTGGCAAGCGAGGATTCCGCCGTGTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCTATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 489. 1844 H1 GYAFSSYW G152- W159 490. 1844 H1 GGCTATGCATTTTCTAGTTACTGG -1 491. 1844 H3 ARRETTTVGRYYYAMDY A223- Y239 492. 1844 H3 GCTCGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCTATGGACTAC -1 493. 1844 H2 IWPGDGDT I177- T184 494. 1844 H2 ATTTGGCCCGGGGATGGAGACACC -1 495. 1844 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 496. 1844 CH2 GCTCCTGAGGCTGCAGGAGGACCAAGCGTGTTCCTGTTTCCCCCTAAACCTAAGGACACACTGATGATCTCTC- GGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAG -1 CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACTAAGC- CTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTG TCTGTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- ACTGCCAGCCCCCATCGAGAAGACAATTTCCAAAGCA AAG 497. 1844 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 498. 1844 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCAGGTCTCCC- TGACATGTCTGGTGAAGGGATTTTATCCTTCTGATAT -1 TGCCGTGGAGTGGGAAAGTAATGGCCAGCCAGAAAACAATTACAAGACTACCCCTCCAGTGCTGGATTCTG- ACGGGAGTTTCGCTCTGGTCAGTAAACTGACTGTGGAT

AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGTTGTTCAGTGATGCACGAGGCACTGCACAATCATTACAC- CCAGAAAAGCCTGTCCCTGTCTCCCGGC 499. 9284 Full QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- KSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ -1 GTLVTVSSVEGGSGGSGGSGGSGGVDDIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKR- WIYDTSKLASGVPSRFSGSGSGTDYTLTISSLQPEDAATYYCQ QWSSNPFTFGQGTKLEIKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSV- SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEW- ESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPG 500. 9284 Full CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAAGTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCCGTGGAGGGAGGCTCCGGAGGCTCTGGAGGCAGCGGCGGCT- CCGGCGGCGTGGACGATATCCAGATGACCCAGAGC CCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCGCCTCCTCTAGCGTGTCCTA- CATGAACTGGTATCAGCAGAAGCCAGGCAAGGCCCCCAA GCGGTGGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGATTCAGCGGCTCCGGCTCTGGCA- CCGACTATACCCTGACAATCTCCTCTCTGCAGCCCGAGG ATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTTTGGCCAGGGCACAAAGCTGGAG- ATCAAGGCCGCCGAGCCCAAGTCTAGCGACAAGACCCA CACATGCCCACCTTGTCCGGCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAA- AGGATACCCTGATGATCAGCCGCACCCCTGAGGTGACAT GCGTGGTGGTGAGCGTGTCCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTG- CACAATGCCAAGACAAAGCCTCGGGAGGAGCAGTAC AACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAA- GTGCAAGGTGTCCAATAAGGCCCTGCCTGCCCCAATCG AGAAGACCATCTCTAAGGCCAAGGGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGAC- GAGCTGACAAAGAACCAGGTGAGCCTGCTGTGCCTGGT GAAGGGCTTCTATCCAAGCGATATCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGA- CCTGGCCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTC TGTATTCCAAGCTGACAGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCAC- GAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCT CTGAGCCCCGGC 501. 9284 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- KSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ Q1-S119 GTLVTVSS 502. 9284 VH CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAAGTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCC 503. 9284 H1 GYTFTRYT G26-T33 504. 9284 H1 GGCTACACCTTCACAAGGTATACC -1 505. 9284 H3 ARYYDDHYSLDY A97-Y108 506. 9284 H3 GCCCGCTACTATGACGATCACTACAGCCTGGATTAT -1 507. 9284 H2 INPSRGYT I51-T58 508. 9284 H2 ATCAACCCTAGCAGGGGCTACACA -1 509. 9284 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKRWIYDTSKLASGVPSRFSGSGSGTDYTLT- ISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIK D138- K243 510. 9284 VL GATATCCAGATGACCCAGAGCCCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCG- CCTCCTCTAGCGTGTCCTACATGAACTGGTATCAGCA -1 GAAGCCAGGCAAGGCCCCCAAGCGGTGGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGAT- TCAGCGGCTCCGGCTCTGGCACCGACTATACCCTGACA ATCTCCTCTCTGCAGCCCGAGGATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTT- TGGCCAGGGCACAAAGCTGGAGATCAAG 511. 9284 L1 SSVSY S164- Y168 512. 9284 L1 TCTAGCGTGTCCTAC -1 513. 9284 L3 QQWSSNPFT Q225- T233 514. 9284 L3 CAGCAGTGGAGCTCCAATCCTTTCACC -1 515. 9284 L2 DTS D186- S188 516. 9284 L2 GATACCAGC -1 517. 9284 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 518. 9284 CH2 GCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAAAGGATACCCTGATGATCAGCC- GCACCCCTGAGGTGACATGCGTGGTGGTGAGCGTGT -1 CCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACAAAG- CCTCGGGAGGAGCAGTACAACTCTACCTATAGAGTGG TGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGTCCAATAAG- GCCCTGCCTGCCCCAATCGAGAAGACCATCTCTAAGG CCAAG 519. 9284 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476 520. 9284 CH3 GGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGACGAGCTGACAAAGAACCAGGTGAGCC- TGCTGTGCCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCT- GATGGCAGCTTCTTTCTGTATTCCAAGCTGACAGTGGAT AAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCACGAGGCCCTGCACAATCACTACAC- CCAGAAGTCCCTGTCTCTGAGCCCCGGC 521. 9285 Full QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- KSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ -1 GTLVTVSSVEGGSGGSGGSGGSGGVDDIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKR- LIYDTSKLASGVPSRFSGSGSGTDYTLTISSLQPEDAATYYCQQ WSSNPFTFGQGTKLEIKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVS- HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWE- SNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPG 522. 9285 Full CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAAGTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCCGTGGAGGGAGGCTCCGGAGGCTCTGGAGGCAGCGGCGGCT- CCGGCGGCGTGGACGATATCCAGATGACCCAGAGC CCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCGCCTCCTCTAGCGTGTCCTA- CATGAACTGGTATCAGCAGAAGCCAGGCAAGGCCCCCAA GCGGCTGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGATTCAGCGGCTCCGGCTCTGGCA- CCGACTATACCCTGACAATCTCCTCTCTGCAGCCCGAGG ATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTTTGGCCAGGGCACAAAGCTGGAG- ATCAAGGCCGCCGAGCCCAAGTCTAGCGACAAGACCCA CACATGCCCACCTTGTCCGGCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAA- AGGATACCCTGATGATCAGCCGCACCCCTGAGGTGACAT GCGTGGTGGTGAGCGTGTCCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTG- CACAATGCCAAGACAAAGCCTCGGGAGGAGCAGTAC AACTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAA- GTGCAAGGTGTCCAATAAGGCCCTGCCTGCCCCAATCG AGAAGACCATCTCTAAGGCCAAGGGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGAC- GAGCTGACAAAGAACCAGGTGAGCCTGCTGTGCCTGGT GAAGGGCTTCTATCCAAGCGATATCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGA- CCTGGCCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTC TGTATTCCAAGCTGACAGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCAC- GAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCT CTGAGCCCCGGC 523. 9285 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- KSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ Q1-S119 GTLVTVSS 524. 9285 VH CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAAGTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCC 525. 9285 H1 GYTFTRYT G26-T33 526. 9285 H1 GGCTACACCTTCACAAGGTATACC -1 527. 9285 H3 ARYYDDHYSLDY A97-Y108 528. 9285 H3 GCCCGCTACTATGACGATCACTACAGCCTGGATTAT -1 529. 9285 H2 INPSRGYT I51-T58 530. 9285 H2 ATCAACCCTAGCAGGGGCTACACA -1 D138- 531. 9285 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDYTLT- ISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIK K243 532. 9285 VL GATATCCAGATGACCCAGAGCCCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCG- CCTCCTCTAGCGTGTCCTACATGAACTGGTATCAGCA -1 GAAGCCAGGCAAGGCCCCCAAGCGGCTGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGAT- TCAGCGGCTCCGGCTCTGGCACCGACTATACCCTGACA ATCTCCTCTCTGCAGCCCGAGGATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTT- TGGCCAGGGCACAAAGCTGGAGATCAAG 533. 9285 L1 SSVSY S164- Y168 534. 9285 L1 TCTAGCGTGTCCTAC -1 535. 9285 L3 QQWSSNPFT Q225- T233 536. 9285 L3 CAGCAGTGGAGCTCCAATCCTTTCACC -1 537. 9285 L2 DTS D186- S188 538. 9285 L2 GATACCAGC -1 539. 9285 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 540. 9285 CH2 GCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAAAGGATACCCTGATGATCAGCC- GCACCCCTGAGGTGACATGCGTGGTGGTGAGCGTGT -1 CCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACAAAG- CCTCGGGAGGAGCAGTACAACTCTACCTATAGAGTGG TGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGTCCAATAAG- GCCCTGCCTGCCCCAATCGAGAAGACCATCTCTAAGG CCAAG 541. 9285 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476 542. 9285 CH3 GGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGACGAGCTGACAAAGAACCAGGTGAGCC- TGCTGTGCCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCT- GATGGCAGCTTCTTTCTGTATTCCAAGCTGACAGTGGAT AAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCACGAGGCCCTGCACAATCACTACAC- CCAGAAGTCCCTGTCTCTGAGCCCCGGC 543. 9286 Full QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- NSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ -1 GTLVTVSSVEGGSGGSGGSGGSGGVDDIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKR- WIYDTSKLASGVPSRFSGSGSGTDYTLTISSLQPEDAATYYCQ QWSSNPFTFGQGTKLEIKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSV-

SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEW- ESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPG 544. 9286 Full CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAACTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCCGTGGAGGGAGGCTCCGGAGGCTCTGGAGGCAGCGGCGGCT- CCGGCGGCGTGGACGATATCCAGATGACCCAGAGC CCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCGCCTCCTCTAGCGTGTCCTA- CATGAACTGGTATCAGCAGAAGCCAGGCAAGGCCCCCAA GCGGTGGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGATTCAGCGGCTCCGGCTCTGGCA- CCGACTATACCCTGACAATCTCCTCTCTGCAGCCCGAGG ATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTTTGGCCAGGGCACAAAGCTGGAG- ATCAAGGCCGCCGAGCCCAAGTCTAGCGACAAGACCCA CACATGCCCACCTTGTCCGGCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAA- AGGATACCCTGATGATCAGCCGCACCCCTGAGGTGACAT GCGTGGTGGTGAGCGTGTCCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTG- CACAATGCCAAGACAAAGCCTCGGGAGGAGCAGTAC AATTCTACCTATAGAGTGGTGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAA- GTGCAAGGTGTCCAATAAGGCCCTGCCTGCCCCAATCG AGAAGACCATCTCTAAGGCCAAGGGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGAC- GAGCTGACAAAGAACCAGGTGAGCCTGCTGTGCCTGGT GAAGGGCTTCTATCCAAGCGATATCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGA- CCTGGCCCCCTGTGCTGGACTCTGATGGCAGCTTCTTTC TGTATTCCAAGCTGACAGTGGATAAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCAC- GAGGCCCTGCACAATCACTACACCCAGAAGTCCCTGTCT CTGAGCCCCGGC 545. 9286 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- NSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ Q1-S119 GTLVTVSS 546. 9286 VH CAGGTGCAGCTGGTGCAGAGCGGAGGAGGAGTGGTGCAGCCAGGCCGGTCCCTGAGACTGTCTTGTAAGGCCA- GCGGCTACACCTTCACAAGGTATACCATGCACTGG -1 GTGCGCCAGGCACCAGGCAAGGGACTGGAGTGGATCGGCTACATCAACCCTAGCAGGGGCTACACAAACTA- TAATCAGAAGGTGAAGGGCCGCTTCACCATCTCCACA GACAACTCTAAGAATACCGCCTACCTGCAGATGGACTCCCTGAGGGCCGAGGATACAGGCGTGTATTTTTG- CGCCCGCTACTATGACGATCACTACAGCCTGGATTATTG GGGCCAGGGCACCCTGGTGACAGTGAGCTCC 547. 9286 H1 GYTFTRYT G26-T33 548. 9286 H1 GGCTACACCTTCACAAGGTATACC -1 549. 9286 H3 ARYYDDHYSLDY A97-Y108 550. 9286 H3 GCCCGCTACTATGACGATCACTACAGCCTGGATTAT -1 551. 9286 H2 INPSRGYT I51-T58 552. 9286 H2 ATCAACCCTAGCAGGGGCTACACA -1 553. 9286 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKRWIYDTSKLASGVPSRFSGSGSGTDYTLT- ISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIK D138- K243 554. 9286 VL GATATCCAGATGACCCAGAGCCCTTCTAGCCTGTCCGCCTCTGTGGGCGACAGGGTGACCATCACATGTAGCG- CCTCCTCTAGCGTGTCCTACATGAACTGGTATCAGCA -1 GAAGCCAGGCAAGGCCCCCAAGCGGTGGATCTACGATACCAGCAAGCTGGCCTCCGGCGTGCCATCTAGAT- TCAGCGGCTCCGGCTCTGGCACCGACTATACCCTGACA ATCTCCTCTCTGCAGCCCGAGGATGCCGCCACATACTATTGCCAGCAGTGGAGCTCCAATCCTTTCACCTT- TGGCCAGGGCACAAAGCTGGAGATCAAG 555. 9286 L1 SSVSY S164- Y168 556. 9286 L1 TCTAGCGTGTCCTAC -1 557. 9286 L3 QQWSSNPFT Q225- T233 558. 9286 L3 CAGCAGTGGAGCTCCAATCCTTTCACC -1 559. 9286 L2 DTS D186- S188 560. 9286 L2 GATACCAGC -1 561. 9286 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 562. 9286 CH2 GCGCCAGAGGCAGCAGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAGCCAAAGGATACCCTGATGATCAGCC- GCACCCCTGAGGTGACATGCGTGGTGGTGAGCGTGT -1 CCCACGAGGACCCAGAGGTGAAGTTTAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACAAAG- CCTCGGGAGGAGCAGTACAATTCTACCTATAGAGTGG TGAGCGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAGGTGTCCAATAAG- GCCCTGCCTGCCCCAATCGAGAAGACCATCTCTAAGG CCAAG 563. 9286 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476 564. 9286 CH3 GGCCAGCCTCGCGAACCTCAGGTGTACGTGCTGCCTCCATCTAGAGACGAGCTGACAAAGAACCAGGTGAGCC- TGCTGTGCCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TCGCCGTGGAGTGGGAGTCCAATGGCCAGCCCGAGAACAATTACCTGACCTGGCCCCCTGTGCTGGACTCT- GATGGCAGCTTCTTTCTGTATTCCAAGCTGACAGTGGAT AAGTCTAGGTGGCAGCAGGGCAACGTGTTTTCTTGCAGCGTGATGCACGAGGCCCTGCACAATCACTACAC- CCAGAAGTCCCTGTCTCTGAGCCCCGGC 565. 7239 Full DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIKGGGGS -1 GGGGSGGGGSQVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDGDTNYA- QKFQGRATLTADESTSTAYMELSSLRSEDTAVYYCARRE TTTVGRYYYAMDYWGQGTTVTVSSEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV- VVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPS- DIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG 566. 7239 Full GATATTCAGCTGACCCAGAGCCCAAGCTCCCTGTCTGCCAGTGTGGGGGATAGGGCTACAATCACTTGCCGCG- CATCACAGAGCGTGGACTATGAGGGCGATTCCTATC -1 TGAACTGGTACCAGCAGAAGCCAGGGAAAGCACCCAAGCTGCTGATCTACGACGCCTCTAATCTGGTGAGT- GGCATTCCCTCAAGGTTCTCCGGATCTGGCAGTGGGAC TGACTTTACCCTGACAATCTCTAGTGTGCAGCCCGAGGATGCCGCTACCTACTATTGCCAGCAGTCTACAG- AAGACCCTTGGACTTTCGGATGTGGCACCAAACTGgAGA TTAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTCCAGCTGGTGCAGAGC- GGAGCAGAGGTCAAGAAACCCGGAGCCAGCGTG AAAATTTCCTGCAAGGCCTCTGGCTATGCTTTCTCAAGCTACTGGATGAACTGGGTGAGGCAGGCACCAGG- ACAGTGTCTGGAATGGATCGGACAGATTTGGCCTGGGG ACGGAGATACCAATTATGCTCAGAAGTTTCAGGGACGCGCAACTCTGACCGCCGATGAGTCAACAAGCACT- GCATACATGGAGCTGTCCTCTCTGCGCTCCGAAGACACA GCCGTGTACTATTGCGCACGGAGAGAAACCACAACTGTGGGCCGATACTATTACGCAATGGATTACTGGGG- CCAGGGGACCACAGTCACTGTGAGTTCAGAGCCTAAAA GCTCCGACAAGACCCACACATGCCCACCTTGTCCGGCGCCAGAAGCAGCCGGAGGGCCTAGCGTGTTCCTG- TTTCCACCCAAGCCAAAAGATACCCTGATGATCAGCCG GACTCCTGAGGTCACCTGCGTGGTCGTGTCCGTGTCTCACGAGGACCCAGAAGTCAAATTCAACTGGTATG- TGGATGGCGTCGAAGTGCATAATGCTAAGACAAAACCC CGAGAGGAACAGTATAACTCCACCTACCGGGTCGTGTCTGTCCTGACAGTGCTGCATCAGGACTGGCTGAA- CGGCAAGGAGTACAAGTGCAAAGTGAGCAACAAGGCC CTGCCCGCCCCAATCGAAAAGACCATTTCCAAGGCCAAAGGGCAGCCTCGCGAACCTCAGGTCTACGTGTA- CCCTCCATCTAGGGATGAACTGACAAAAAACCAGGTCA GTCTGACTTGTCTGGTGAAGGGCTTCTACCCAAGCGACATTGCCGTGGAGTGGGAATCCAATGGCCAGCCC- GAGAACAATTACAAGACTACCCCCCCTGTGCTGGACAG CGATGGGTCCTTCGCTCTGGTCAGTAAACTGACAGTGGATAAGTCAAGATGGCAGCAGGGAAATGTCTTTA- GTTGTTCAGTGATGCACGAGGCACTGCACAACCACTAC ACCCAGAAGTCACTGTCCCTGTCACCCGGC 567. 7239 VL DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIK D1-K111 568. 7239 VL GATATTCAGCTGACCCAGAGCCCAAGCTCCCTGTCTGCCAGTGTGGGGGATAGGGCTACAATCACTTGCCGCG- CATCACAGAGCGTGGACTATGAGGGCGATTCCTATC TGAACTGGTACCAGCAGAAGCCAGGGAAAGCACCCAAGCTGCTGATCTACGACGCCTCTAATCTGGTGAGT- GGCATTCCCTCAAGGTTCTCCGGATCTGGCAGTGGGAC TGACTTTACCCTGACAATCTCTAGTGTGCAGCCCGAGGATGCCGCTACCTACTATTGCCAGCAGTCTACAG- AAGACCCTTGGACTTTCGGATGTGGCACCAAACTGGAGA TTAAG -1 569. 7239 L1 QSVDYEGDSY Q27-Y36 570. 7239 L1 CAGAGCGTGGACTATGAGGGCGATTCCTAT -1 571. 7239 L3 QQSTEDPWT Q93- T101 572. 7239 L3 CAGCAGTCTACAGAAGACCCTTGGACT -1 573. 7239 L2 DAS D54-S56 574. 7239 L2 GACGCCTCT -1 575. 7239 VH QVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDGDTNYAQKFQGRATLTAD- ESTSTAYMELSSLRSEDTAVYYCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 576. 7239 VH CAGGTCCAGCTGGTGCAGAGCGGAGCAGAGGTCAAGAAACCCGGAGCCAGCGTGAAAATTTCCTGCAAGGCCT- CTGGCTATGCTTTCTCAAGCTACTGGATGAACTGG -1 GTGAGGCAGGCACCAGGACAGTGTCTGGAATGGATCGGACAGATTTGGCCTGGGGACGGAGATACCAATTA- TGCTCAGAAGTTTCAGGGACGCGCAACTCTGACCGCC GATGAGTCAACAAGCACTGCATACATGGAGCTGTCCTCTCTGCGCTCCGAAGACACAGCCGTGTACTATTG- CGCACGGAGAGAAACCACAACTGTGGGCCGATACTATT ACGCAATGGATTACTGGGGCCAGGGGACCACAGTCACTGTGAGTTCA 577. 7239 H1 GYAFSSYW G152- W159 578. 7239 H1 GGCTATGCTTTCTCAAGCTACTGG -1 579. 7239 H3 ARRETTTVGRYYYAMDY A223- Y239 580. 7239 H3 GCACGGAGAGAAACCACAACTGTGGGCCGATACTATTACGCAATGGATTAC -1 581. 7239 H2 IWPGDGDT I177- T184 582. 7239 H2 ATTTGGCCTGGGGACGGAGATACC -1 583. 7239 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A266- K375 584. 7239 CH2 GCGCCAGAAGCAGCCGGAGGGCCTAGCGTGTTCCTGTTTCCACCCAAGCCAAAAGATACCCTGATGATCAGCC- GGACTCCTGAGGTCACCTGCGTGGTCGTGTCCGTGT -1 CTCACGAGGACCCAGAAGTCAAATTCAACTGGTATGTGGATGGCGTCGAAGTGCATAATGCTAAGACAAAA- CCCCGAGAGGAACAGTATAACTCCACCTACCGGGTCGT GTCTGTCCTGACAGTGCTGCATCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAAGTGAGCAACAAGG- CCCTGCCCGCCCCAATCGAAAAGACCATTTCCAAGGCC AAA 585. 7239 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G376- G481 586. 7239 CH3 GGGCAGCCTCGCGAACCTCAGGTCTACGTGTACCCTCCATCTAGGGATGAACTGACAAAAAACCAGGTCAGTC- TGACTTGTCTGGTGAAGGGCTTCTACCCAAGCGACAT -1 TGCCGTGGAGTGGGAATCCAATGGCCAGCCCGAGAACAATTACAAGACTACCCCCCCTGTGCTGGACAGCG- ATGGGTCCTTCGCTCTGGTCAGTAAACTGACAGTGGAT AAGTCAAGATGGCAGCAGGGAAATGTCTTTAGTTGTTCAGTGATGCACGAGGCACTGCACAACCACTACAC- CCAGAAGTCACTGTCCCTGTCACCCGGC 587. 9288 Full QVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDGDTNYAQKFQGRATLTAD- ESTSTAYMELSSLRSEDTAVYYCARRETTTVGRYYYAM -1 DYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ- SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKP- REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFA- LVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 588. 9288 Full CAGGTCCAGCTGGTGCAGAGCGGAGCAGAGGTGAAGAAGCCAGGAGCCAGCGTGAAGATCTCCTGCAAGGCCT- CTGGCTATGCCTTCAGCTCCTACTGGATGAACTGG -1 GTGCGGCAGGCACCTGGACAGTGTCTGGAGTGGATCGGACAGATCTGGCCAGGCGACGGCGATACAAATTA-

TGCCCAGAAGTTTCAGGGCAGAGCCACACTGACCGCC GACGAGAGCACATCCACCGCCTACATGGAGCTGTCTAGCCTGAGGAGCGAGGATACCGCCGTGTACTATTG- CGCAAGGAGAGAGACCACAACCGTGGGCCGCTACTAT TACGCCATGGACTATTGGGGCCAGGGCACAACCGTGACAGTGTCCTCTGCTAGCACCAAGGGACCTTCCGT- GTTCCCACTGGCACCAAGCTCCAAGTCTACAAGCGGAG GAACCGCCGCCCTGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGTGACCGTGTCTTGGAACAGCGGG- GCCCTGACCAGCGGAGTGCACACCTTTCCTGCCGTGCT GCAGTCTAGCGGCCTGTATTCCCTGTCCTCTGTGGTCACAGTGCCAAGCTCCTCTCTGGGCACACAGACCT- ACATCTGCAACGTGAATCACAAGCCATCCAATACCAAGGT CGACAAGAAGGTGGAGCCCAAGTCTTGTGATAAGACACACACCTGCCCACCTTGTCCGGCGCCAGAGGCAG- CAGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAGCCT AAGGACACACTGATGATCTCCAGGACACCAGAGGTGACCTGCGTGGTGGTGTCCGTGTCTCACGAGGACCC- CGAGGTGAAGTTCAACTGGTACGTGGATGGCGTGGAG GTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTATAACTCTACATACCGCGTGGTGAGCGTGCTGAC- CGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTAC AAGTGCAAGGTGAGCAATAAGGCCCTGCCCGCCCCTATCGAGAAGACCATCTCCAAGGCCAAGGGCCAGCC- TCGCGAACCACAGGTGTACGTGTACCCTCCATCTAGAG ACGAGCTGACAAAGAACCAGGTGAGCCTGACCTGTCTGGTGAAGGGCTTTTATCCCAGCGATATCGCCGTG- GAGTGGGAGTCCAATGGCCAGCCTGAGAACAATTACAA GACAACCCCCCCTGTGCTGGACTCCGATGGCTCTTTCGCCCTGGTGTCCAAGCTGACCGTGGACAAGTCTC- GGTGGCAGCAGGGCAACGTGTTCAGCTGTTCCGTGATGC ACGAGGCACTGCACAATCACTACACCCAGAAGTCACTGTCACTGTCCCCAGGC 589. 9288 VH QVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDGDTNYAQKFQGRATLTAD- ESTSTAYMELSSLRSEDTAVYYCARRETTTVGRYYYAM Q1-S124 DYWGQGTTVTVSS 590. 9288 VH CAGGTCCAGCTGGTGCAGAGCGGAGCAGAGGTGAAGAAGCCAGGAGCCAGCGTGAAGATCTCCTGCAAGGCCT- CTGGCTATGCCTTCAGCTCCTACTGGATGAACTGG -1 GTGCGGCAGGCACCTGGACAGTGTCTGGAGTGGATCGGACAGATCTGGCCAGGCGACGGCGATACAAATTA- TGCCCAGAAGTTTCAGGGCAGAGCCACACTGACCGCC GACGAGAGCACATCCACCGCCTACATGGAGCTGTCTAGCCTGAGGAGCGAGGATACCGCCGTGTACTATTG- CGCAAGGAGAGAGACCACAACCGTGGGCCGCTACTAT TACGCCATGGACTATTGGGGCCAGGGCACAACCGTGACAGTGTCCTCT 591. 9288 H1 GYAFSSYW G26-W33 592. 9288 H1 GGCTATGCCTTCAGCTCCTACTGG -1 593. 9288 H3 ARRETTTVGRYYYAMDY A97-Y113 594. 9288 H3 GCAAGGAGAGAGACCACAACCGTGGGCCGCTACTATTACGCCATGGACTAT -1 595. 9288 H2 IWPGDGDT I51-T58 596. 9288 H2 ATCTGGCCAGGCGACGGCGATACA -1 597. 9288 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A125- V222 598. 9288 CH1 GCTAGCACCAAGGGACCTTCCGTGTTCCCACTGGCACCAAGCTCCAAGTCTACAAGCGGAGGAACCGCCGCCC- TGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCG -1 TGACCGTGTCTTGGAACAGCGGGGCCCTGACCAGCGGAGTGCACACCTTTCCTGCCGTGCTGCAGTCTAGC- GGCCTGTATTCCCTGTCCTCTGTGGTCACAGTGCCAAGC TCCTCTCTGGGCACACAGACCTACATCTGCAACGTGAATCACAAGCCATCCAATACCAAGGTCGACAAGAA- GGTG 599. 9288 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A238- K347 600. 9288 CH2 GCGCCAGAGGCAGCAGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAGCCTAAGGACACACTGATGATCTCCA- GGACACCAGAGGTGACCTGCGTGGTGGTGTCCGTGT -1 CTCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGATGGCGTGGAGGTGCACAATGCCAAGACCAAG- CCCAGGGAGGAGCAGTATAACTCTACATACCGCGTGG TGAGCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGAGCAATAAG- GCCCTGCCCGCCCCTATCGAGAAGACCATCTCCAAGG CCAAG 601. 9288 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G348- G453 602. 9288 CH3 GGCCAGCCTCGCGAACCACAGGTGTACGTGTACCCTCCATCTAGAGACGAGCTGACAAAGAACCAGGTGAGCC- TGACCTGTCTGGTGAAGGGCTTTTATCCCAGCGATA -1 TCGCCGTGGAGTGGGAGTCCAATGGCCAGCCTGAGAACAATTACAAGACAACCCCCCCTGTGCTGGACTCC- GATGGCTCTTTCGCCCTGGTGTCCAAGCTGACCGTGGA CAAGTCTCGGTGGCAGCAGGGCAACGTGTTCAGCTGTTCCGTGATGCACGAGGCACTGCACAATCACTACA- CCCAGAAGTCACTGTCACTGTCCCCAGGC 603. 9289 Full DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIKRTVAAP -1 SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA- DYEKHKVYACEVTHQGL5SPVTKSFNRGEC 604. 9289 Full GATATTCAGCTGACCCAGTCTCCAAGCTCCCTGAGCGCCTCCGTGGGCGATAGGGCCACCATCACATGCAGAG- CCTCTCAGAGCGTGGACTACGAGGGCGATTCCTACCT -1 GAACTGGTATCAGCAGAAGCCAGGCAAGGCCCCCAAGCTGCTGATCTATGACGCCTCCAATCTGGTGTCTG- GCATCCCCAGCCGGTTCTCCGGCTCTGGCAGCGGAACA GACTTTACCCTGACAATCTCTAGCGTGCAGCCTGAGGATGCCGCCACCTACTATTGCCAGCAGAGCACCGA- GGACCCATGGACATTCGGCTGTGGCACCAAGCTGGAGA TCAAGAGGACAGTGGCGGCGCCCAGCGTGTTCATCTTTCCCCCTTCCGATGAGCAGCTGAAGTCCGGCACC- GCCTCTGTGGTGTGCCTGCTGAACAACTTCTACCCCCGG GAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGTCCGGCAATTCTCAGGAGAGCGTGACAGAGCA- GGACTCCAAGGATTCTACCTATAGCCTGTCCTCTACCC TGACACTGTCCAAGGCCGATTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACCAGGGGCTGTCA- TCACCAGTCACCAAATCATTCAATAGGGGCGAGTGC 605. 9289 VL DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIK D1-K111 606. 9289 VL GATATTCAGCTGACCCAGTCTCCAAGCTCCCTGAGCGCCTCCGTGGGCGATAGGGCCACCATCACATGCAGAG- CCTCTCAGAGCGTGGACTACGAGGGCGATTCCTACCT -1 GAACTGGTATCAGCAGAAGCCAGGCAAGGCCCCCAAGCTGCTGATCTATGACGCCTCCAATCTGGTGTCTG- GCATCCCCAGCCGGTTCTCCGGCTCTGGCAGCGGAACA GACTTTACCCTGACAATCTCTAGCGTGCAGCCTGAGGATGCCGCCACCTACTATTGCCAGCAGAGCACCGA- GGACCCATGGACATTCGGCTGTGGCACCAAGCTGGAGA TCAAG 607. 9289 L1 QSVDYEGDSY Q27-Y36 608. 9289 L1 CAGAGCGTGGACTACGAGGGCGATTCCTAC -1 Q93-T101 609. 9289 L3 QQSTEDPWT 610. 9289 L3 CAGCAGAGCACCGAGGACCCATGGACA -1 611. 9289 L2 DAS D54-S56 612. 9289 L2 GACGCCTCC -1 613. 9289 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R112- C218 614. 9289 CL AGGACAGTGGCGGCGCCCAGCGTGTTCATCTTTCCCCCTTCCGATGAGCAGCTGAAGTCCGGCACCGCCTCTG- TGGTGTGCCTGCTGAACAACTTCTACCCCCGGGAGGC -1 CAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGTCCGGCAATTCTCAGGAGAGCGTGACAGAGCAGGACT- CCAAGGATTCTACCTATAGCCTGTCCTCTACCCTGACA CTGTCCAAGGCCGATTACGAGAAGCACAAGGTGTATGCCTGTGAGGTCACCCACCAGGGGCTGTCATCACC- AGTCACCAAATCATTCAATAGGGGCGAGTGC 615. 5239 Full QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKDRFTISRD- NSKNTAFLQMDSLRPEDTGVYFCARYYDDHYCLDYWGQ -1 GTPVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS5GLY- SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN- STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTV- DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 616. 5239 Full CAGGTCCAGCTGGTCCAGTCCGGAGGAGGAGTGGTCCAGCCAGGACGGTCACTGAGACTGAGCTGCAAGGCTT- CCGGGTACACTTTCACCCGATATACCATGCACTGG -1 GTGCGGCAGGCACCAGGGAAAGGACTGGAATGGATCGGGTACATTAACCCTAGCAGGGGATACACAAACTA- TAATCAGAAGGTGAAAGACAGGTTCACTATCTCTCGC GATAACAGTAAGAATACCGCCTTTCTGCAGATGGACAGCCTGCGCCCCGAGGATACAGGCGTGTATTTCTG- CGCTCGATACTATGACGATCACTACTGTCTGGACTATTG GGGCCAGGGGACTCCAGTCACCGTGAGCTCCGCATCAACTAAGGGACCCAGCGTGTTTCCACTGGCCCCCT- CTAGTAAATCCACATCTGGAGGAACTGCAGCTCTGGGA TGCCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTGAGCTGGAACTCCGGAGCCCTGACTTCCGGCGT- CCATACCTTTCCCGCTGTGCTGCAGTCAAGCGGGCTGTA CTCTCTGTCCTCTGTGGTCACAGTGCCTAGTTCAAGCCTGGGAACACAGACTTATATCTGCAACGTGAATC- ACAAGCCTAGCAATACTAAAGTCGACAAGAAAGTGGAAC CAAAGAGCTGTGATAAAACCCATACATGCCCCCCTTGTCCTGCACCAGAGGCAGCAGGAGGACCAAGCGTG- TTCCTGTTTCCACCCAAGCCTAAAGACACCCTGATGATT AGCCGGACCCCTGAAGTGACATGTGTGGTCGTGAGTGTGTCACACGAGGACCCAGAAGTCAAGTTCAACTG- GTACGTGGATGGCGTCGAGGTGCATAATGCCAAGACA AAACCTAGAGAGGAACAGTACAATTCCACCTATAGGGTCGTGTCTGTCCTGACAGTGCTGCACCAGGATTG- GCTGAACGGGAAAGAGTATAAGTGCAAAGTGTCCAATA AGGCTCTGCCCGCACCTATCGAGAAAACCATTTCTAAGGCTAAAGGCCAGCCTAGGGAACCACAGGTCTAC- GTGTATCCTCCATCTCGCGACGAGCTGACAAAGAACCA GGTCAGTCTGACTTGTCTGGTGAAAGGATTTTACCCAAGCGATATTGCCGTGGAGTGGGAATCCAATGGCC- AGCCCGAAAACAATTATAAGACCACACCCCCTGTGCTGG ACTCTGATGGCAGTTTCGCACTGGTCAGTAAGCTGACTGTGGACAAATCAAGATGGCAGCAGGGGAACGTC- TTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCA TTACACCCAGAAGTCTCTGAGTCTGTCACCCGGC 617. 5239 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKDRFTISRD- NSKNTAFLQMDSLRPEDTGVYFCARYYDDHYCLDYWGQ Q1-S119 GTPVTVSS 618. 5239 VH CAGGTCCAGCTGGTCCAGTCCGGAGGAGGAGTGGTCCAGCCAGGACGGTCACTGAGACTGAGCTGCAAGGCTT- CCGGGTACACTTTCACCCGATATACCATGCACTGG -1 GTGCGGCAGGCACCAGGGAAAGGACTGGAATGGATCGGGTACATTAACCCTAGCAGGGGATACACAAACTA- TAATCAGAAGGTGAAAGACAGGTTCACTATCTCTCGC GATAACAGTAAGAATACCGCCTTTCTGCAGATGGACAGCCTGCGCCCCGAGGATACAGGCGTGTATTTCTG- CGCTCGATACTATGACGATCACTACTGTCTGGACTATTG GGGCCAGGGGACTCCAGTCACCGTGAGCTCC 619. 5239 H1 GYTFTRYT G26-T33 620. 5239 H1 GGGTACACTTTCACCCGATATACC -1 621. 5239 H3 ARYYDDHYCLDY A97-Y108 622. 5239 H3 GCTCGATACTATGACGATCACTACTGTCTGGACTAT -1 623. 5239 H2 INPSRGYT I51-T58 624. 5239 H2 ATTAACCCTAGCAGGGGATACACA -1 625. 5239 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A120- V217 626. 5239 CH1 GCATCAACTAAGGGACCCAGCGTGTTTCCACTGGCCCCCTCTAGTAAATCCACATCTGGAGGAACTGCAGCTC- TGGGATGCCTGGTGAAGGATTACTTCCCAGAGCCCGT -1 CACCGTGAGCTGGAACTCCGGAGCCCTGACTTCCGGCGTCCATACCTTTCCCGCTGTGCTGCAGTCAAGCG- GGCTGTACTCTCTGTCCTCTGTGGTCACAGTGCCTAGTTC AAGCCTGGGAACACAGACTTATATCTGCAACGTGAATCACAAGCCTAGCAATACTAAAGTCGACAAGAAAG- TG 627. 5239 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A233- K342 628. 5239 CH2 GCACCAGAGGCAGCAGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAGCCTAAAGACACCCTGATGATTAGCC- GGACCCCTGAAGTGACATGTGTGGTCGTGAGTGTGT -1 CACACGAGGACCCAGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAGACAAAA- CCTAGAGAGGAACAGTACAATTCCACCTATAGGGTCG TGTCTGTCCTGACAGTGCTGCACCAGGATTGGCTGAACGGGAAAGAGTATAAGTGCAAAGTGTCCAATAAG- GCTCTGCCCGCACCTATCGAGAAAACCATTTCTAAGGCT AAA 629. 5239 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G343- G448 630. 5239 CH3 GGCCAGCCTAGGGAACCACAGGTCTACGTGTATCCTCCATCTCGCGACGAGCTGACAAAGAACCAGGTCAGTC- TGACTTGTCTGGTGAAAGGATTTTACCCAAGCGATAT TGCCGTGGAGTGGGAATCCAATGGCCAGCCCGAAAACAATTATAAGACCACACCCCCTGTGCTGGACTCTG- ATGGCAGTTTCGCACTGGTCAGTAAGCTGACTGTGGAC AAATCAAGATGGCAGCAGGGGAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAGTCTCTGAGTCTGTCACCCGGC -1

631. 2304 Full QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKDRFTISRD- NSKNTAFLQMDSLRPEDTGVYFCARYYDDHYCLDYWGQ -1 GTPVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY- SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTC PPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN- STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTV- DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 632. 2304 Full CAGGTCCAGCTGGTGCAGAGCGGAGGAGGAGTGGTCCAGCCAGGACGGTCTCTGAGACTGAGTTGCAAGGCAT- CAGGGTACACTTTCACCCGATATACCATGCACTGG -1 GTGCGGCAGGCACCAGGGAAAGGACTGGAATGGATCGGGTACATTAACCCTTCCAGGGGATACACAAACTA- TAATCAGAAGGTGAAAGACAGGTTCACTATCAGCCGC GATAACTCCAAGAATACCGCTTTTCTGCAGATGGACTCTCTGCGCCCCGAGGATACAGGCGTGTATTTCTG- CGCACGATACTATGACGATCACTACTGTCTGGACTATTGG GGCCAGGGGACTCCAGTCACCGTGAGCTCCGCCTCTACTAAGGGACCCAGTGTGTTTCCACTGGCTCCCTC- TAGTAAATCCACATCTGGAGGAACTGCAGCTCTGGGATG CCTGGTGAAGGATTACTTCCCAGAGCCCGTCACCGTGAGTTGGAACTCAGGAGCTCTGACTAGCGGCGTCC- ATACCTTTCCCGCAGTGCTGCAGTCAAGCGGGCTGTACA GCCTGTCCTCTGTGGTCACAGTGCCTAGTTCAAGCCTGGGAACACAGACTTATATCTGCAACGTGAATCAC- AAGCCTTCTAATACTAAAGTCGACAAGAAAGTGGAACCA AAGAGTTGTGATAAAACCCATACATGCCCACCTTGTCCTGCACCAGAGCTGCTGGGAGGACCAAGCGTGTT- CCTGTTTCCACCCAAGCCTAAAGACACCCTGATGATTAG CCGGACCCCTGAAGTCACATGTGTGGTCGTGGACGTGAGCCACGAGGACCCCGAAGTCAAGTTCAACTGGT- ACGTGGATGGCGTCGAGGTGCATAATGCCAAGACAAA ACCTAGAGAGGAACAGTACAATTCAACCTATAGGGTCGTGAGCGTCCTGACAGTGCTGCACCAGGACTGGC- TGAACGGGAAGGAGTATAAGTGCAAAGTGTCCAATAA GGCACTGCCCGCCCCTATCGAGAAAACCATTTCTAAGGCAAAAGGCCAGCCTAGGGAACCACAGGTCTACG- TGTATCCTCCAAGCCGCGACGAGCTGACAAAGAACCAG GTCTCCCTGACTTGTCTGGTGAAAGGATTTTACCCAAGTGATATTGCTGTGGAGTGGGAATCAAATGGCCA- GCCCGAAAACAATTATAAGACCACACCCCCTGTGCTGGA CAGCGATGGCTCCTTCGCCCTGGTCTCCAAGCTGACTGTGGATAAATCTAGATGGCAGCAGGGGAACGTCT- TTAGTTGTTCAGTGATGCATGAGGCTCTGCACAATCATT ACACCCAGAAGAGCCTGTCCCTGTCTCCCGGCAAA 633. 2304 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKDRFTISRD- NSKNTAFLQMDSLRPEDTGVYFCARYYDDHYCLDYWGQ Q1-S119 GTPVTVSS 634. 2304 VH CAGGTCCAGCTGGTGCAGAGCGGAGGAGGAGTGGTCCAGCCAGGACGGTCTCTGAGACTGAGTTGCAAGGCAT- CAGGGTACACTTTCACCCGATATACCATGCACTGG -1 GTGCGGCAGGCACCAGGGAAAGGACTGGAATGGATCGGGTACATTAACCCTTCCAGGGGATACACAAACTA- TAATCAGAAGGTGAAAGACAGGTTCACTATCAGCCGC GATAACTCCAAGAATACCGCTTTTCTGCAGATGGACTCTCTGCGCCCCGAGGATACAGGCGTGTATTTCTG- CGCACGATACTATGACGATCACTACTGTCTGGACTATTGG GGCCAGGGGACTCCAGTCACCGTGAGCTCC 635. 2304 H1 GYTFTRYT G26-T33 636. 2304 H1 GGGTACACTTTCACCCGATATACC -1 637. 2304 H3 ARYYDDHYCLDY A97-Y108 638. 2304 H3 GCACGATACTATGACGATCACTACTGTCTGGACTAT -1 639. 2304 H2 INPSRGYT I51-T58 640. 2304 H2 ATTAACCCTTCCAGGGGATACACA -1 641. 2304 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A120- V217 642. 2304 CH1 GCCTCTACTAAGGGACCCAGTGTGTTTCCACTGGCTCCCTCTAGTAAATCCACATCTGGAGGAACTGCAGCTC- TGGGATGCCTGGTGAAGGATTACTTCCCAGAGCCCGT -1 CACCGTGAGTTGGAACTCAGGAGCTCTGACTAGCGGCGTCCATACCTTTCCCGCAGTGCTGCAGTCAAGCG- GGCTGTACAGCCTGTCCTCTGTGGTCACAGTGCCTAGTT CAAGCCTGGGAACACAGACTTATATCTGCAACGTGAATCACAAGCCTTCTAATACTAAAGTCGACAAGAAA- GTG 643. 2304 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A233- K342 644. 2304 CH2 GCACCAGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAGCCTAAAGACACCCTGATGATTAGCC- GGACCCCTGAAGTCACATGTGTGGTCGTGGACGTGA -1 GCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAGACAAAA- CCTAGAGAGGAACAGTACAATTCAACCTATAGGGTCG TGAGCGTCCTGACAGTGCTGCACCAGGACTGGCTGAACGGGAAGGAGTATAAGTGCAAAGTGTCCAATAAG- GCACTGCCCGCCCCTATCGAGAAAACCATTTCTAAGGC AAAA 645. 2304 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G343- G448 646. 2304 CH3 GGCCAGCCTAGGGAACCACAGGTCTACGTGTATCCTCCAAGCCGCGACGAGCTGACAAAGAACCAGGTCTCCC- TGACTTGTCTGGTGAAAGGATTTTACCCAAGTGATAT -1 TGCTGTGGAGTGGGAATCAAATGGCCAGCCCGAAAACAATTATAAGACCACACCCCCTGTGCTGGACAGCG- ATGGCTCCTTCGCCCTGGTCTCCAAGCTGACTGTGGATA AATCTAGATGGCAGCAGGGGAACGTCTTTAGTTGTTCAGTGATGCATGAGGCTCTGCACAATCATTACACC- CAGAAGAGCCTGTCCCTGTCTCCCGGC 647. 3537 Full QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDN- SKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGT -1 LVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL- SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST- YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQP REPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDK- SRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 648. 3537 Full CAGGTCCAGCTGAAGCAGAGCGGACCAGGACTGGTGCAGCCTTCACAGAGCCTGTCCATCACCTGCACAGTGT- CCGGATTCTCTCTGACTAACTACGGAGTCCACTGGGT -1 GCGACAGAGTCCAGGAAAAGGCCTGGAGTGGCTGGGCGTGATCTGGAGCGGAGGGAACACCGACTATAATA- CCCCTTTTACAAGTCGGCTGTCAATTAACAAGGATAA CTCTAAGAGTCAGGTGTTCTTTAAGATGAACAGCCTGCAGTCCAATGACACCGCTATCTACTATTGCGCTA- GAGCACTGACATACTATGATTACGAGTTCGCATATTGGGG GCAGGGAACTCTGGTCACCGTGTCTGCCGCTAGTACAAAGGGACCAAGCGTGTTTCCACTGGCACCAAGCT- CCAAATCAACAAGCGGAGGCACTGCAGCCCTGGGATGT CTGGTGAAGGACTACTTCCCAGAGCCCGTCACTGTGTCATGGAACAGCGGCGCACTGACTTCCGGGGTCCA- TACCTTTCCTGCCGTGCTGCAGTCTAGTGGCCTGTACTCT CTGTCAAGCGTGGTCACAGTGCCATCCTCTAGTCTGGGGACTCAGACCTATATCTGCAACGTGAATCACAA- GCCTTCCAATACTAAAGTCGACAAGAAAGTGGAACCAAA GTCTTGTGATAAAACACATACTTGCCCCCCTTGTCCTGCACCAGAGCTGCTGGGGGGACCATCCGTGTTCC- TGTTTCCACCCAAGCCCAAAGACACCCTGATGATTTCCCG CACCCCAGAAGTCACATGCGTGGTCGTGGACGTGTCTCACGAGGACCCCGAAGTCAAGTTCAACTGGTACG- TGGATGGCGTCGAGGTGCATAATGCTAAGACAAAACCA CGGGAGGAACAGTACAATAGTACATATAGAGTCGTGTCAGTGCTGACCGTCCTGCACCAGGACTGGCTGAA- CGGCAAGGAGTATAAGTGCAAAGTGAGCAATAAGGCC CTGCCCGCTCCTATCGAGAAAACCATTAGCAAGGCCAAAGGGCAGCCTAGGGAACCACAGGTCTACGTGCT- GCCTCCATCACGCGACGAGCTGACAAAGAACCAGGTCA GCCTGCTGTGTCTGGTGAAAGGGTTCTATCCCTCTGATATCGCTGTGGAGTGGGAAAGTAATGGACAGCCT- GAAAACAATTACCTGACTTGGCCCCCTGTGCTGGACTCC GATGGATCTTTCTTTCTGTATAGCAAGCTGACCGTGGATAAATCCAGGTGGCAGCAGGGCAACGTCTTTTC- CTGTTCTGTGATGCATGAGGCCCTGCACAATCATTACACC CAGAAGAGTCTGTCACTGAGCCCTGGCAAA 649. 3537 VH QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDN- SKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGT Q1-A119 LVTVSA 650. 3537 VH CAGGTCCAGCTGAAGCAGAGCGGACCAGGACTGGTGCAGCCTTCACAGAGCCTGTCCATCACCTGCACAGTGT- CCGGATTCTCTCTGACTAACTACGGAGTCCACTGGGT -1 GCGACAGAGTCCAGGAAAAGGCCTGGAGTGGCTGGGCGTGATCTGGAGCGGAGGGAACACCGACTATAATA- CCCCTTTTACAAGTCGGCTGTCAATTAACAAGGATAA CTCTAAGAGTCAGGTGTTCTTTAAGATGAACAGCCTGCAGTCCAATGACACCGCTATCTACTATTGCGCTA- GAGCACTGACATACTATGATTACGAGTTCGCATATTGGGG GCAGGGAACTCTGGTCACCGTGTCTGCC 651. 3537 H1 GFSLTNYG G26-G33 652. 3537 H1 GGATTCTCTCTGACTAACTACGGA -1 653. 3537 H3 ARALTYYDYEFAY A96-Y108 654. 3537 H3 GCTAGAGCACTGACATACTATGATTACGAGTTCGCATAT -1 655. 3537 H2 IWSGGNT I51-T57 656. 3537 H2 ATCTGGAGCGGAGGGAACACC -1 657. 3537 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A120- V217 658. 3537 CH1 GCTAGTACAAAGGGACCAAGCGTGTTTCCACTGGCACCAAGCTCCAAATCAACAAGCGGAGGCACTGCAGCCC- TGGGATGTCTGGTGAAGGACTACTTCCCAGAGCCCG -1 TCACTGTGTCATGGAACAGCGGCGCACTGACTTCCGGGGTCCATACCTTTCCTGCCGTGCTGCAGTCTAGT- GGCCTGTACTCTCTGTCAAGCGTGGTCACAGTGCCATCCT CTAGTCTGGGGACTCAGACCTATATCTGCAACGTGAATCACAAGCCTTCCAATACTAAAGTCGACAAGAAA- GTG 659. 3537 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A233- K342 660. 3537 CH2 GCACCAGAGCTGCTGGGGGGACCATCCGTGTTCCTGTTTCCACCCAAGCCCAAAGACACCCTGATGATTTCCC- GCACCCCAGAAGTCACATGCGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCTAAGACAAAAC- CACGGGAGGAACAGTACAATAGTACATATAGAGTCGT GTCAGTGCTGACCGTCCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAAGTGAGCAATAAGG- CCCTGCCCGCTCCTATCGAGAAAACCATTAGCAAGGCC AAA 661. 3537 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G343- G448 662. 3537 CH3 GGGCAGCCTAGGGAACCACAGGTCTACGTGCTGCCTCCATCACGCGACGAGCTGACAAAGAACCAGGTCAGCC- TGCTGTGTCTGGTGAAAGGGTTCTATCCCTCTGATA -1 TCGCTGTGGAGTGGGAAAGTAATGGACAGCCTGAAAACAATTACCTGACTTGGCCCCCTGTGCTGGACTCC- GATGGATCTTTCTTTCTGTATAGCAAGCTGACCGTGGAT AAATCCAGGTGGCAGCAGGGCAACGTCTTTTCCTGTTCTGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAGAGTCTGTCACTGAGCCCTGGC 663. 3299 Full QVQLVQSGAEVKKPGASVKVSCKASGYTFRSSYISWVRQAPGQGLEWMGWIYAGTGSPSYNQKLQGRVTMTTD- TSTSTAYMELRSLRSDDTAVYYCARHRDYYSNSLTYWG -1 QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL- YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY- NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLT- VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 664. 3299 Full CAGGTCCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAACCCGGGGCTTCTGTCAAGGTGAGTTGCAAAGCTT- CAGGCTACACATTCCGAAGCTCCTATATCAGCTGGG -1 TGCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGGCTGGATCTACGCTGGCACAGGGAGTCCCTCATAT- AACCAGAAGCTGCAGGGGCGGGTGACTATGACCACA GACACTAGCACCTCCACAGCATACATGGAACTGAGGTCCCTGCGCTCTGACGATACCGCTGTGTACTATTG- CGCACGGCACAGAGATTACTATTCTAATAGTCTGACCTAT TGGGGACAGGGCACCCTGGTCACAGTGTCTAGTGCTAGCACAAAGGGGCCTTCCGTGTTTCCACTGGCACC- CTCAAGCAAATCAACTAGCGGAGGAACCGCAGCTCTGG GATGTCTGGTGAAGGACTACTTCCCAGAGCCCGTCACAGTGAGTTGGAACTCAGGGGCACTGACCAGCGGA- GTCCATACATTTCCTGCCGTGCTGCAGTCCTCTGGGCT GTACTCCCTGAGTTCAGTGGTCACAGTGCCAAGCTCCTCTCTGGGAACTCAGACCTATATCTGCAACGTGA- ATCACAAGCCATCCAATACTAAAGTCGACAAGAAAGTGG AACCCAAGTCTTGTGATAAAACACATACTTGCCCACCTTGTCCTGCACCAGAGCTGCTGGGAGGACCATCC- GTGTTCCTGTTTCCACCCAAGCCTAAAGACACTCTGATGA TTAGCAGGACACCCGAAGTCACTTGCGTGGTCGTGGACGTGAGCCACGAGGACCCCGAAGTCAAGTTCAAC- TGGTACGTGGATGGCGTCGAGGTGCATAATGCTAAGA CCAAACCCAGAGAGGAACAGTACAACTCTACTTATAGGGTCGTGAGTGTCCTGACCGTGCTGCACCAGGAC- TGGCTGAACGGCAAGGAGTATAAGTGCAAAGTGTCCAA TAAGGCACTGCCCGCCCCTATCGAGAAAACCATTTCTAAGGCAAAAGGGCAGCCTCGCGAACCACAGGTCT- ACGTGCTGCCTCCAAGTCGAGACGAGCTGACAAAGAAC CAGGTCAGCCTGCTGTGTCTGGTGAAAGGATTCTATCCTTCCGATATCGCCGTGGAGTGGGAATCTAATGG- CCAGCCAGAGAACAATTACCTGACTTGGCCCCCTGTGCT GGACAGCGATGGCAGCTTCTTCCTGTATTCAAAGCTGACCGTGGATAAAAGCCGGTGGCAGCAGGGCAACG- TCTTTTCCTGTTCTGTGATGCATGAAGCCCTGCACAATC ATTACACCCAGAAGAGTCTGTCACTGAGCCCCGGCAAA 665. 3299 VH QVQLVQSGAEVKKPGASVKVSCKASGYTRSSYISWVRQAPGQGLEWMGWIYAGTGSPSYNQKLQGRVTMTTDT- STSTAYMELRSLRSDDTAVYYCARHRDYYSNSLTYWG Q1-S120 QGTLVTVSS 666. 3299 VH CAGGTCCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAACCCGGGGCTTCTGTCAAGGTGAGTTGCAAAGCTT- CAGGCTACACATTCCGAAGCTCCTATATCAGCTGGG -1

TGCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGGCTGGATCTACGCTGGCACAGGGAGTCCCTCATAT- AACCAGAAGCTGCAGGGGCGGGTGACTATGACCACA GACACTAGCACCTCCACAGCATACATGGAACTGAGGTCCCTGCGCTCTGACGATACCGCTGTGTACTATTG- CGCACGGCACAGAGATTACTATTCTAATAGTCTGACCTAT TGGGGACAGGGCACCCTGGTCACAGTGTCTAGT 667. 3299 H1 GYTFRSSY G26-Y33 668. 3299 H1 GGCTACACATTCCGAAGCTCCTAT -1 669. 3299 H3 ARHRDYYSNSLTY A97-Y109 670. 3299 H3 GCACGGCACAGAGATTACTATTCTAATAGTCTGACCTAT -1 671. 3299 H2 IYAGTGSP I51-P58 672. 3299 H2 ATCTACGCTGGCACAGGGAGTCCC -1 673. 3299 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS5GLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A121- V218 674. 3299 CH1 GCTAGCACAAAGGGGCCTTCCGTGTTTCCACTGGCACCCTCAAGCAAATCAACTAGCGGAGGAACCGCAGCTC- TGGGATGTCTGGTGAAGGACTACTTCCCAGAGCCCG -1 TCACAGTGAGTTGGAACTCAGGGGCACTGACCAGCGGAGTCCATACATTTCCTGCCGTGCTGCAGTCCTCT- GGGCTGTACTCCCTGAGTTCAGTGGTCACAGTGCCAAGC TCCTCTCTGGGAACTCAGACCTATATCTGCAACGTGAATCACAAGCCATCCAATACTAAAGTCGACAAGAA- AGTG 675. 3299 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A234- K343 676. 3299 CH2 GCACCAGAGCTGCTGGGAGGACCATCCGTGTTCCTGTTTCCACCCAAGCCTAAAGACACTCTGATGATTAGCA- GGACACCCGAAGTCACTTGCGTGGTCGTGGACGTGA -1 GCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCTAAGACCAAA- CCCAGAGAGGAACAGTACAACTCTACTTATAGGGTCGT GAGTGTCCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTATAAGTGCAAAGTGTCCAATAAGG- CACTGCCCGCCCCTATCGAGAAAACCATTTCTAAGGCA AAA 677. 3299 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G344- G449 678. 3299 CH3 GGGCAGCCTCGCGAACCACAGGTCTACGTGCTGCCTCCAAGTCGAGACGAGCTGACAAAGAACCAGGTCAGCC- TGCTGTGTCTGGTGAAAGGATTCTATCCTTCCGATA -1 TCGCCGTGGAGTGGGAATCTAATGGCCAGCCAGAGAACAATTACCTGACTTGGCCCCCTGTGCTGGACAGC- GATGGCAGCTTCTTCCTGTATTCAAAGCTGACCGTGGAT AAAAGCCGGTGGCAGCAGGGCAACGTCTTTTCCTGTTCTGTGATGCATGAAGCCCTGCACAATCATTACAC- CCAGAAGAGTCTGTCACTGAGCCCCGGC 679. 8071 Full QIVLTQSPAIM55SPGEKVTMSCSATSSVTYMYWYQQKPGSSPKPWIFRTSNLASGVPTRFSGSGSGTSYSLT- ISSMEAEDAATYYCQHYHIYPRTFGGGTKLELKRTVAAPSVFIF -1 PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH- KVYACEVTHQGLSSPVTKSFNRGEC 680. 8071 Full CAGATCGTCCTGACTCAGAGCCCCGCTATCATGTCCTCATCCCCTGGCGAGAAGGTCACTATGTCATGCTCCG- CTACTTCCTCCGTGACATACATGTATTGGTACCAGCAG -1 AAGCCAGGCAGCTCCCCCAAACCTTGGATCTTCCGGACTTCCAACCTGGCAAGCGGGGTGCCCACCAGATT- TTCAGGCAGCGGGTCCGGAACCTCTTATAGTCTGACAAT TTCTAGTATGGAGGCCGAAGACGCCGCTACCTACTATTGCCAGCACTACCATATCTACCCAAGAACATTCG- GAGGCGGAACTAAACTGGAGCTGAAACGAACCGTGGCG GCGCCCAGTGTCTTCATTTTTCCCCCTAGCGACGAACAGCTGAAGTCTGGGACAGCCAGTGTGGTCTGTCT- GCTGAACAACTTCTACCCTAGAGAGGCTAAAGTGCAGTG GAAGGTCGATAACGCACTGCAGTCCGGAAATTCTCAGGAGAGTGTGACTGAACAGGACTCAAAAGATAGCA- CCTATTCCCTGTCAAGCACACTGACTCTGAGCAAGGCC GACTACGAGAAGCATAAAGTGTATGCTTGTGAAGTCACCCACCAGGGGCTGAGTTCACCAGTCACAAAATC- ATTCAACAGAGGGGAGTGC 681. 8071 VL QIVLTQSPAIM55SPGEKVTMSCSATSSVTYMYWYQQKPGSSPKPWIFRTSNLASGVPTRFSGSGSGTSYSLT- ISSMEAEDAATYYCQHYHIYPRTFGGGTKLELK Q1-K106 682. 8071 VL CAGATCGTCCTGACTCAGAGCCCCGCTATCATGTCCTCATCCCCTGGCGAGAAGGTCACTATGTCATGCTCCG- CTACTTCCTCCGTGACATACATGTATTGGTACCAGCAG -1 AAGCCAGGCAGCTCCCCCAAACCTTGGATCTTCCGGACTTCCAACCTGGCAAGCGGGGTGCCCACCAGATT- TTCAGGCAGCGGGTCCGGAACCTCTTATAGTCTGACAAT TTCTAGTATGGAGGCCGAAGACGCCGCTACCTACTATTGCCAGCACTACCATATCTACCCAAGAACATTCG- GAGGCGGAACTAAACTGGAGCTGAAA 683. 8071 L1 SSVTY S27-Y31 684. 8071 L1 TCCTCCGTGACATAC -1 685. 8071 L3 QHYHIYPRT Q88-T96 686. 8071 L3 CAGCACTACCATATCTACCCAAGAACA -1 687. 8071 L2 RTS R49-S51 688. 8071 L2 CGGACTTCC -1 689. 8071 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R107- C213 690. 8071 CL CGAACCGTGGCGGCGCCCAGTGTCTTCATTTTTCCCCCTAGCGACGAACAGCTGAAGTCTGGGACAGCCAGTG- TGGTCTGTCTGCTGAACAACTTCTACCCTAGAGAGGC -1 TAAAGTGCAGTGGAAGGTCGATAACGCACTGCAGTCCGGAAATTCTCAGGAGAGTGTGACTGAACAGGACT- CAAAAGATAGCACCTATTCCCTGTCAAGCACACTGACT CTGAGCAAGGCCGACTACGAGAAGCATAAAGTGTATGCTTGTGAAGTCACCCACCAGGGGCTGAGTTCACC- AGTCACAAAATCATTCAACAGAGGGGAGTGC 691. 1109 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSGGGGSDIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPG- QGLEWIGYINPSRGYTNYNQKFKDKATLTTDK5SSTAYMQ LSSLTSEDSAVYYCARYYDDHYCLDYWGQGTTLTVSSVEGGSGGSGGSGGSGGVDDIQLTQSPAIMSASPG- EKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGV PYRFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTKLELKHHHHHH 692. 1109 Full GATATTCAGCTGACACAGTCTCCAGCTAGTCTGGCAGTGAGCCTGGGCCAGCGGGCTACTATCAGCTGCAAGG- CAAGCCAGTCCGTCGACTACGATGGGGACAGCTATC -1 TGAACTGGTACCAGCAGATCCCCGGACAGCCCCCTAAACTGCTGATCTACGACGCCTCAAATCTGGTGAGC- GGCATCCCACCCAGATTCTCTGGAAGTGGCTCAGGGACC GATTTTACACTGAACATTCACCCCGTGGAGAAGGTCGACGCCGCTACCTACCATTGCCAGCAGTCCACTGA- GGACCCCTGGACCTTCGGAGGAGGAACAAAGCTGGAAA TCAAAGGCGGAGGAGGCAGTGGAGGAGGAGGGAGCGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAACTGGTGAGACCTGGAAGCTCCGTCA AGATTTCCTGTAAAGCATCTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGACTGGAGTGGATCGGACAGATTTGGCCTGGGGA TGGAGACACCAACTACAATGGAAAGTTCAAAGGCAAGGCTACCCTGACAGCAGACGAATCAAGCTCCACAG- CTTACATGCAGCTGTCTAGTCTGGCATCAGAGGATAGC GCCGTGTATTTTTGCGCTCGGAGAGAAACCACAACTGTCGGCCGCTACTATTACGCCATGGACTACTGGGG- CCAGGGGACCACAGTGACAGTCTCAAGCGGCGGGGGA GGCTCCGATATCAAGCTGCAGCAGTCTGGAGCAGAGCTGGCTCGACCAGGAGCCAGTGTGAAGATGTCATG- TAAAACCAGCGGCTATACTTTCACCAGGTACACAATGC ACTGGGTGAAACAGCGCCCAGGACAGGGCCTGGAATGGATCGGATACATTAACCCCTCCAGGGGCTATACC- AACTACAATCAGAAGTTCAAGGATAAAGCCACTCTGAC TACCGACAAGTCCTCTAGTACCGCTTATATGCAGCTGTCAAGCCTGACATCCGAGGACTCTGCAGTGTATT- ACTGCGCCCGCTATTACGACGATCATTATTGTCTGGATTA CTGGGGGCAGGGAACAACTCTGACTGTGTCCTCTGTCGAAGGGGGAAGTGGAGGGTCAGGAGGCAGCGGAG- GCAGCGGAGGGGTGGACGATATCCAGCTGACCCAG TCCCCTGCCATTATGAGCGCTTCCCCAGGCGAGAAGGTGACAATGACTTGCAGGGCTAGTTCAAGCGTCTC- TTATATGAATTGGTATCAGCAGAAGTCTGGCACTAGTCC TAAACGATGGATCTATGACACCTCCAAAGTGGCATCTGGGGTCCCATACCGGTTCTCTGGCAGTGGGTCAG- GAACTAGCTATTCCCTGACCATTTCCTCTATGGAGGCAG AAGATGCAGCCACCTATTACTGTCAGCAGTGGAGTTCAAATCCCCTGACATTTGGCGCCGGGACTAAGCTG- GAGCTGAAACACCATCACCATCACCAT 693. 1109 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 694. 1109 VL GATATTCAGCTGACACAGTCTCCAGCTAGTCTGGCAGTGAGCCTGGGCCAGCGGGCTACTATCAGCTGCAAGG- CAAGCCAGTCCGTCGACTACGATGGGGACAGCTATC -1 TGAACTGGTACCAGCAGATCCCCGGACAGCCCCCTAAACTGCTGATCTACGACGCCTCAAATCTGGTGAGC- GGCATCCCACCCAGATTCTCTGGAAGTGGCTCAGGGACC GATTTTACACTGAACATTCACCCCGTGGAGAAGGTCGACGCCGCTACCTACCATTGCCAGCAGTCCACTGA- GGACCCCTGGACCTTCGGAGGAGGAACAAAGCTGGAAA TCAAA 695. 1109 L1 QSVDYDGDSY Q27-Y36 696. 1109 L1 CAGTCCGTCGACTACGATGGGGACAGCTAT -1 697. 1109 L3 QQSTEDPWT Q93- T101 698. 1109 L3 CAGCAGTCCACTGAGGACCCCTGGACC -1 699. 1109 L2 DAS D54-S56 700. 1109 L2 GACGCCTCA -1 701. 1109 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS 5250 702. 1109 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAACTGGTGAGACCTGGAAGCTCCGTCAAGATTTCCTGTAAAGCAT- CTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGACTGGAGTGGATCGGACAGATTTGGCCTGGGGATGGAGACACCAACTAC- AATGGAAAGTTCAAAGGCAAGGCTACCCTGACAGCA GACGAATCAAGCTCCACAGCTTACATGCAGCTGTCTAGTCTGGCATCAGAGGATAGCGCCGTGTATTTTTG- CGCTCGGAGAGAAACCACAACTGTCGGCCGCTACTATTA CGCCATGGACTACTGGGGCCAGGGGACCACAGTGACAGTCTCAAGC 703. 1109 H1 GYAFSSYW G152- W159 704. 1109 H1 GGCTATGCCTTTTCTAGTTACTGG -1 705. 1109 H3 ARRETTTVGRYYYAMDY A223- Y239 706. 1109 H3 GCTCGGAGAGAAACCACAACTGTCGGCCGCTACTATTACGCCATGGACTAC -1 707. 1109 H2 IWPGDGDT I177- T184 708. 1109 H2 ATTTGGCCTGGGGATGGAGACACC -1 709. 1109 VH DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG D256- TTLTVSS S374 710. 1109 VH GATATCAAGCTGCAGCAGTCTGGAGCAGAGCTGGCTCGACCAGGAGCCAGTGTGAAGATGTCATGTAAAACCA- GCGGCTATACTTTCACCAGGTACACAATGCACTGGG -1 TGAAACAGCGCCCAGGACAGGGCCTGGAATGGATCGGATACATTAACCCCTCCAGGGGCTATACCAACTAC- AATCAGAAGTTCAAGGATAAAGCCACTCTGACTACCGA CAAGTCCTCTAGTACCGCTTATATGCAGCTGTCAAGCCTGACATCCGAGGACTCTGCAGTGTATTACTGCG- CCCGCTATTACGACGATCATTATTGTCTGGATTACTGGGG GCAGGGAACAACTCTGACTGTGTCCTCT 711. 1109 H1 GYTFTRYT G281- T288 712. 1109 H1 GGCTATACTTTCACCAGGTACACA -1 713. 1109 H3 ARYYDDHYCLDY A352- Y363 714. 1109 H3 GCCCGCTATTACGACGATCATTATTGTCTGGATTAC -1 715. 1109 H2 INPSRGYT I306- T313 716. 1109 H2 ATTAACCCCTCCAGGGGCTATACC -1 717. 1109 VL DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFSGSGSGTSYSLT- ISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK D393- K498 718. 1109 VL GATATCCAGCTGACCCAGTCCCCTGCCATTATGAGCGCTTCCCCAGGCGAGAAGGTGACAATGACTTGCAGGG- CTAGTTCAAGCGTCTCTTATATGAATTGGTATCAGCA -1 GAAGTCTGGCACTAGTCCTAAACGATGGATCTATGACACCTCCAAAGTGGCATCTGGGGTCCCATACCGGT- TCTCTGGCAGTGGGTCAGGAACTAGCTATTCCCTGACCA TTTCCTCTATGGAGGCAGAAGATGCAGCCACCTATTACTGTCAGCAGTGGAGTTCAAATCCCCTGACATTT-

GGCGCCGGGACTAAGCTGGAGCTGAAA 719. 1109 L1 SSVSY S419- Y423 720. 1109 L1 TCAAGCGTCTCTTAT -1 721. 1109 L3 QQWSSNPLT Q480- T488 722. 1109 L3 CAGCAGTGGAGTTCAAATCCCCTGACA -1 723. 1109 L2 DTS D441- S443 724. 1109 L2 GACACCTCC -1 725. 2170 Full DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG -1 TTLTVSSVEGGSGGSGGSGGSGGVDDIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRW- IYDTSKVASGVPYRFSGSGSGTSYSLTISSMEAEDAATYYCQ QWSSNPLTFGAGTKLELKAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV- SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEW- ESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGK 726. 2170 Full GACATCAAACTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCCAGTGTGAAAATGTCATGCAAGACAA- GCGGCTACACCTTCACACGGTATACTATGCACTGG -1 GTGAAACAGAGACCCGGCCAGGGGCTGGAATGGATCGGATATATTAACCCTTCCCGAGGCTACACCAACTA- TAATCAGAAGTTTAAAGACAAGGCCACCCTGACCACAG ATAAGAGCTCCTCTACAGCTTACATGCAGCTGAGTTCACTGACTAGTGAGGACTCAGCTGTGTACTATTGC- GCAAGGTACTATGACGATCATTACTGTCTGGATTATTGGG GACAGGGCACTACCCTGACTGTCAGCTCCGTGGAAGGAGGGAGCGGAGGCTCCGGAGGATCTGGCGGGAGT- GGAGGCGTGGACGATATCCAGCTGACCCAGTCCCCA GCAATTATGTCCGCCTCTCCCGGCGAGAAAGTGACTATGACCTGCCGCGCCTCTAGTTCAGTGAGCTACAT- GAACTGGTATCAGCAGAAATCAGGCACCAGCCCCAAGAG ATGGATCTACGACACATCCAAGGTCGCTTCTGGGGTGCCTTATAGGTTCAGTGGGTCAGGAAGCGGCACTT- CCTACTCTCTGACCATTAGCTCCATGGAGGCAGAAGATG CCGCTACATACTATTGTCAGCAGTGGTCTAGTAATCCACTGACATTTGGGGCCGGAACTAAACTGGAGCTG- AAGGCAGCCGAACCCAAATCAAGCGACAAGACACACAC TTGCCCACCTTGTCCAGCACCAGAACTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGG- ATACACTGATGATCAGCCGGACCCCTGAGGTCACATGCG TGGTCGTGGACGTGAGCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCAT- AATGCCAAAACCAAGCCTAGGGAGGAACAGTACAATA GTACTTATCGCGTCGTGTCAGTCCTGACCGTGCTGCATCAGGATTGGCTGAACGGGAAGGAGTACAAATGC- AAGGTGTCCAACAAGGCCCTGCCTGCTCCAATCGAGAA GACCATTTCTAAAGCAAAGGGCCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCCCGGGACGAGC- TGACCAAAAACCAGGTCTCTCTGACATGTCTGGTGAAGG GGTTTTATCCATCTGATATTGCTGTGGAGTGGGAAAGTAATGGACAGCCCGAGAACAATTACAAGACAACT- CCCCCTGTGCTGGACTCCGATGGATCTTTCGCTCTGGTC AGCAAACTGACAGTGGACAAGTCCAGATGGCAGCAGGGCAACGTCTTTAGTTGTTCAGTGATGCACGAGGC- ACTGCACAATCATTACACTCAGAAAAGCCTGTCCCTGT CTCCCGGCAAG 727. 2170 VH DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG D1-S119 TTLTVSS 728. 2170 VH GACATCAAACTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCCAGTGTGAAAATGTCATGCAAGACAA- GCGGCTACACCTTCACACGGTATACTATGCACTGG -1 GTGAAACAGAGACCCGGCCAGGGGCTGGAATGGATCGGATATATTAACCCTTCCCGAGGCTACACCAACTA- TAATCAGAAGTTTAAAGACAAGGCCACCCTGACCACAG ATAAGAGCTCCTCTACAGCTTACATGCAGCTGAGTTCACTGACTAGTGAGGACTCAGCTGTGTACTATTGC- GCAAGGTACTATGACGATCATTACTGTCTGGATTATTGGG GACAGGGCACTACCCTGACTGTCAGCTCC 729. 2170 H1 GYTFTRYT G26-T33 730. 2170 H1 GGCTACACCTTCACACGGTATACT -1 731. 2170 H3 ARYYDDHYCLDY A97-Y108 732. 2170 H3 GCAAGGTACTATGACGATCATTACTGTCTGGATTAT -1 733. 2170 H2 INPSRGYT I51-T58 734. 2170 H2 ATTAACCCTTCCCGAGGCTACACC -1 735. 2170 VL DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFSGSGSGTSYSLT- ISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK D138- K243 736. 2170 VL GATATCCAGCTGACCCAGTCCCCAGCAATTATGTCCGCCTCTCCCGGCGAGAAAGTGACTATGACCTGCCGCG- CCTCTAGTTCAGTGAGCTACATGAACTGGTATCAGCA -1 GAAATCAGGCACCAGCCCCAAGAGATGGATCTACGACACATCCAAGGTCGCTTCTGGGGTGCCTTATAGGT- TCAGTGGGTCAGGAAGCGGCACTTCCTACTCTCTGACCA TTAGCTCCATGGAGGCAGAAGATGCCGCTACATACTATTGTCAGCAGTGGTCTAGTAATCCACTGACATTT- GGGGCCGGAACTAAACTGGAGCTGAAG S164- 737. 2170 L1 SSVSY Y168 738. 2170 L1 AGTTCAGTGAGCTAC -1 739. 2170 L3 QQWSSNPLT Q225- T233 740. 2170 L3 CAGCAGTGGTCTAGTAATCCACTGACA -1 741. 2170 L2 DTS D186- S188 742. 2170 L2 GACACATCC -1 743. 2170 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 744. 2170 CH2 GCACCAGAACTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTGATGATCAGCC- GGACCCCTGAGGTCACATGCGTGGTCGTGGACGTGA -1 GCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCATAATGCCAAAACCAAG- CCTAGGGAGGAACAGTACAATAGTACTTATCGCGTCG TGTCAGTCCTGACCGTGCTGCATCAGGATTGGCTGAACGGGAAGGAGTACAAATGCAAGGTGTCCAACAAG- GCCCTGCCTGCTCCAATCGAGAAGACCATTTCTAAAGC AAAG 745. 2170 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476 746. 2170 CH3 GGCCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCCCGGGACGAGCTGACCAAAAACCAGGTCTCTC- TGACATGTCTGGTGAAGGGGTTTTATCCATCTGATAT -1 TGCTGTGGAGTGGGAAAGTAATGGACAGCCCGAGAACAATTACAAGACAACTCCCCCTGTGCTGGACTCCG- ATGGATCTTTCGCTCTGGTCAGCAAACTGACAGTGGAC AAGTCCAGATGGCAGCAGGGCAACGTCTTTAGTTGTTCAGTGATGCACGAGGCACTGCACAATCATTACAC- TCAGAAAAGCCTGTCCCTGTCTCCCGGC 747. 1890 Full DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG -1 TTLTVSSVEGGSGGSGGSGGSGGVDDIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRW- IYDTSKVASGVPYRFSGSGSGTSYSLTISSMEAEDAATYYCQ QWSSNPLTFGAGTKLELKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV- SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVL HQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVE- WESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK 748. 1890 Full GACATCAAACTGCAGCAGAGCGGAGCAGAGCTGGCTCGACCAGGAGCCAGTGTGAAAATGTCATGCAAGACCA- GCGGCTACACATTCACTCGGTATACAATGCACTGG -1 GTGAAGCAGAGACCAGGACAGGGACTGGAATGGATCGGATATATTAACCCTTCCCGAGGCTACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCAACTCTGACCACA GATAAGAGCTCCTCTACCGCCTACATGCAGCTGAGTTCACTGACAAGTGAGGACTCAGCCGTGTACTATTG- CGCTAGGTACTATGACGATCATTACTGTCTGGATTATTGG GGACAGGGCACTACCCTGACTGTCAGCTCCGTGGAAGGAGGGAGCGGAGGCTCCGGAGGATCTGGCGGGAG- TGGAGGCGTGGACGATATCCAGCTGACCCAGTCCCC AGCTATTATGTCCGCATCTCCCGGCGAGAAAGTCACCATGACATGCCGCGCCTCTAGTTCAGTGAGCTACA- TGAACTGGTATCAGCAGAAATCAGGCACTAGCCCCAAGA GATGGATCTACGACACCTCCAAGGTCGCTTCTGGGGTGCCTTATAGGTTCAGTGGGTCAGGAAGCGGCACC- TCCTACTCTCTGACAATTAGCTCCATGGAGGCTGAAGAT GCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAATCCACTGACTTTTGGGGCAGGAACCAAACTGGAGCT- GAAGGCAGCCGAACCCAAATCAAGCGACAAGACTCACA CCTGCCCACCTTGTCCAGCACCAGAAGCTGCAGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAG- GATACACTGATGATCAGCCGGACACCTGAGGTCACTTGC GTGGTCGTGGACGTGAGCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCA- TAATGCCAAAACCAAGCCTAGGGAGGAACAGTACAAT AGTACATATAGAGTCGTGTCAGTGCTGACCGTCCTGCATCAGGATTGGCTGAACGGGAAGGAGTACAAATG- CAAGGTGTCCAACAAGGCACTGCCTGCCCCAATCGAGA AGACCATTTCTAAAGCAAAGGGCCAGCCCCGAGAACCTCAGGTCTATGTGCTGCCTCCATCCCGGGACGAG- CTGACAAAAAACCAGGTCTCTCTGCTGTGTCTGGTGAAG GGGTTCTACCCATCTGATATTGCTGTGGAGTGGGAAAGTAATGGACAGCCCGAGAACAATTATCTGACATG- GCCCCCTGTGCTGGACTCCGATGGATCTTTCTTTCTGTAC AGCAAACTGACTGTGGACAAGTCCAGATGGCAGCAGGGCAACGTCTTTAGTTGTTCAGTGATGCACGAGGC- CCTGCACAATCATTACACCCAGAAAAGCCTGTCCCTGTC TCCCGGCAAG 749. 1890 VH DIKLQQSGAELARPGASVKMSCKTSGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQG D1-S119 TTLTVSS 750. 1890 VH GACATCAAACTGCAGCAGAGCGGAGCAGAGCTGGCTCGACCAGGAGCCAGTGTGAAAATGTCATGCAAGACCA- GCGGCTACACATTCACTCGGTATACAATGCACTGG -1 GTGAAGCAGAGACCAGGACAGGGACTGGAATGGATCGGATATATTAACCCTTCCCGAGGCTACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCAACTCTGACCACA GATAAGAGCTCCTCTACCGCCTACATGCAGCTGAGTTCACTGACAAGTGAGGACTCAGCCGTGTACTATTG- CGCTAGGTACTATGACGATCATTACTGTCTGGATTATTGG GGACAGGGCACTACCCTGACTGTCAGCTCC 751. 1890 H1 GYTFTRYT G26-T33 752. 1890 H1 GGCTACACATTCACTCGGTATACA -1 753. 1890 H3 ARYYDDHYCLDY A97-Y108 754. 1890 H3 GCTAGGTACTATGACGATCATTACTGTCTGGATTAT -1 755. 1890 H2 INPSRGYT I51-T58 756. 1890 H2 ATTAACCCTTCCCGAGGCTACACA -1 757. 1890 VL DIQLTQSPAIMSASPGEKVTMTCRASSSVSYMNWYQQKSGTSPKRWIYDTSKVASGVPYRFSGSGSGTSYSLT- ISSMEAEDAATYYCQQWSSNPLTFGAGTKLELK D138- K243 758. 1890 VL GATATCCAGCTGACCCAGTCCCCAGCTATTATGTCCGCATCTCCCGGCGAGAAAGTCACCATGACATGCCGCG- CCTCTAGTTCAGTGAGCTACATGAACTGGTATCAGCA -1 GAAATCAGGCACTAGCCCCAAGAGATGGATCTACGACACCTCCAAGGTCGCTTCTGGGGTGCCTTATAGGT- TCAGTGGGTCAGGAAGCGGCACCTCCTACTCTCTGACAA TTAGCTCCATGGAGGCTGAAGATGCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAATCCACTGACTTTT- GGGGCAGGAACCAAACTGGAGCTGAAG 759. 1890 L1 SSVSY S164- Y168 760. 1890 L1 AGTTCAGTGAGCTAC -1 761. 1890 L3 QQWSSNPLT Q225- T233 762. 1890 L3 CAGCAGTGGTCTAGTAATCCACTGACT -1 763. 1890 L2 DTS D186- S188 764. 1890 L2 GACACCTCC -1 765. 1890 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A261- K370 766. 1890 CH2 GCACCAGAAGCTGCAGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTGATGATCAGCC- GGACACCTGAGGTCACTTGCGTGGTCGTGGACGTGA -1 GCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCATAATGCCAAAACCAAG- CCTAGGGAGGAACAGTACAATAGTACATATAGAGTCG TGTCAGTGCTGACCGTCCTGCATCAGGATTGGCTGAACGGGAAGGAGTACAAATGCAAGGTGTCCAACAAG- GCACTGCCTGCCCCAATCGAGAAGACCATTTCTAAAGC AAAG 767. 1890 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G371- G476

768. 1890 CH3 GGCCAGCCCCGAGAACCTCAGGTCTATGTGCTGCCTCCATCCCGGGACGAGCTGACAAAAAACCAGGTCTCTC- TGCTGTGTCTGGTGAAGGGGTTCTACCCATCTGATAT -1 TGCTGTGGAGTGGGAAAGTAATGGACAGCCCGAGAACAATTATCTGACATGGCCCCCTGTGCTGGACTCCG- ATGGATCTTTCTTTCTGTACAGCAAACTGACTGTGGACA AGTCCAGATGGCAGCAGGGCAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACACC- CAGAAAAGCCTGTCCCTGTCTCCCGGC 769. 2171 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK- FKDKATLTTDKSSSTAYMQLSSLTSEDSAVYYCARYYDDHY SLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES- NGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 770. 2171 Full CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACCATGACATGCTCAG- CCAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACATCCCCCAAGAGATGGATCTACGACACTTCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACTAGTTATTCACTGACC ATTTCCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACATT- TGGATCTGGCACTAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTCCAGCTGCAGCAGAGCGGAGCTGAGCTGGCA- CGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGC CAGCGGCTACACTTTCACCCGGTATACCATGCATTGGGTGAAACAGAGACCCGGGCAGGGACTGGAATGGA- TCGGGTACATTAATCCTTCCCGAGGATACACAAACTAC AACCAGAAGTTTAAAGACAAGGCTACCCTGACCACAGATAAGAGCTCCTCTACAGCATATATGCAGCTGAG- TTCACTGACTTCTGAGGACAGTGCCGTGTACTATTGCGC TAGGTACTATGACGATCACTACTCCCTGGATTATTGGGGCCAGGGGACTACCCTGACCGTGAGCTCCGCAG- CCGAACCTAAATCTAGTGACAAGACACATACTTGCCCAC CTTGTCCAGCACCAGAGCTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTG- ATGATCTCCCGGACCCCTGAAGTCACATGTGTGGTCGTG GACGTGTCTCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACTAAGCCCAGGGAGGAACAGTACAACTCCACTTATC GCGTCGTGTCTGTCCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGC- AACAAGGCCCTGCCTGCTCCAATCGAGAAGACCATTAG CAAAGCAAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCTCGGGACGAGCTGACCAAAA- ACCAGGTCAGTCTGACATGTCTGGTGAAGGGCTTTTAC CCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTATAAGACAACTCCCCCTGT- GCTGGACTCAGATGGGAGCTTCGCCCTGGTCAGTAAAC TGACTGTGGACAAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCTCTGCAC- AATCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGC AAG 771. 2171 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Qi-N106 772. 2171 VL CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACCATGACATGCTCAG- CCAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACATCCCCCAAGAGATGGATCTACGACACTTCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACTAGTTATTCACTGACC ATTTCCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACATT- TGGATCTGGCACTAAGCTGGAAATTAAT 773. 2171 L1 SSVSY S27-Y31 774. 2171 L1 TCCTCTGTGAGCTAC -1 775. 2171 L3 QQWSSNPFT Q88-T96 776. 2171 L3 CAGCAGTGGAGTTCAAACCCATTCACA -1 777. 2171 L2 DTS D49-S51 778. 2171 L2 GACACTTCC -1 779. 2171 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ Q122- GTTLTVSS S240 780. 2171 VH CAGGTCCAGCTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGCCA- GCGGCTACACTTTCACCCGGTATACCATGCATTGG GTGAAACAGAGACCCGGGCAGGGACTGGAATGGATCGGGTACATTAATCCTTCCCGAGGATACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCTACCCTGACCACA GATAAGAGCTCCTCTACAGCATATATGCAGCTGAGTTCACTGACTTCTGAGGACAGTGCCGTGTACTATTG- CGCTAGGTACTATGACGATCACTACTCCCTGGATTATTGG GGCCAGGGGACTACCCTGACCGTGAGCTCC -1 781. 2171 H1 GYTFTRYT G147- T154 782. 2171 H1 GGCTACACTTTCACCCGGTATACC -1 783. 2171 H3 ARYYDDHYSLDY A218- Y229 784. 2171 H3 GCTAGGTACTATGACGATCACTACTCCCTGGATTAT -1 785. 2171 H2 INPSRGYT I172- T179 786. 2171 H2 ATTAATCCTTCCCGAGGATACACA -1 787. 2171 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 788. 2171 CH2 GCACCAGAGCTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTGATGATCTCCC- GGACCCCTGAAGTCACATGTGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACTAAGC- CCAGGGAGGAACAGTACAACTCCACTTATCGCGTCGTG TCTGTCCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- CCTGCCTGCTCCAATCGAGAAGACCATTAGCAAAGCAA AG 789. 2171 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 790. 2171 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCTCGGGACGAGCTGACCAAAAACCAGGTCAGTC- TGACATGTCTGGTGAAGGGCTTTTACCCAAGCGATAT -1 TGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTATAAGACAACTCCCCCTGTGCTGGACTCAG- ATGGGAGCTTCGCCCTGGTCAGTAAACTGACTGTGGAC AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCTCTGCACAATCATTACAC- CCAGAAATCTCTGAGTCTGTCACCCGGC 791. 3300 Full DIVMTQSPDSLAVSLGERATINCKSSQSVLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGS- GTDFTLTISSLQAEDVAVYYCQSDYSYPYTFGQGTKLEIKRTV -1 AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL- SKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 792. 3300 Full GACATTGTGATGACACAGTCCCCTGACTCTCTGGCTGTGTCCCTGGGCGAGCGAGCAACTATCAATTGCAAGA- GCTCCCAGTCCGTCCTGAACTCTGGGAATCAGAAAAA -1 CTATCTGACCTGGTACCAGCAGAAGCCCGGACAGCCCCCTAAACTGCTGATCTATTGGGCTAGTACCCGGG- AGTCAGGCGTGCCTGACAGATTCAGTGGCTCAGGGAGC GGAACAGATTTTACCCTGACAATTTCTAGTCTGCAGGCAGAAGACGTGGCCGTCTACTATTGCCAGAGTGA- TTACTCATATCCTTACACATTCGGCCAGGGGACTAAGCT GGAGATCAAAAGGACTGTGGCCGCTCCAAGTGTCTTCATTTTTCCACCCTCCGACGAACAGCTGAAGTCCG- GCACAGCCTCTGTGGTCTGTCTGCTGAACAATTTTTATCC ACGCGAGGCCAAGGTGCAGTGGAAAGTCGATAATGCTCTGCAGAGCGGGAACTCCCAGGAGTCTGTGACCG- AACAGGACAGTAAGGATTCAACATACAGCCTGTCAAG CACTCTGACCCTGTCTAAAGCAGATTATGAGAAGCACAAAGTGTACGCCTGCGAAGTCACTCATCAGGGAC- TGTCCTCTCCCGTGACCAAGAGCTTCAACAGAGGCGAAT GT 793. 3300 VL DIVMTQSPDSLAVSLGERATINCKSSQSVLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGS- GTDFTLTISSLQAEDVAVYYCQSDYSYPYTFGQGTKLEIK D1-K113 794. 3300 VL GACATTGTGATGACACAGTCCCCTGACTCTCTGGCTGTGTCCCTGGGCGAGCGAGCAACTATCAATTGCAAGA- GCTCCCAGTCCGTCCTGAACTCTGGGAATCAGAAAAA -1 CTATCTGACCTGGTACCAGCAGAAGCCCGGACAGCCCCCTAAACTGCTGATCTATTGGGCTAGTACCCGGG- AGTCAGGCGTGCCTGACAGATTCAGTGGCTCAGGGAGC GGAACAGATTTTACCCTGACAATTTCTAGTCTGCAGGCAGAAGACGTGGCCGTCTACTATTGCCAGAGTGA- TTACTCATATCCTTACACATTCGGCCAGGGGACTAAGCT GGAGATCAAA 795. 3300 L1 QSVLNSGNQKNY Q27-Y38 796. 3300 L1 CAGTCCGTCCTGAACTCTGGGAATCAGAAAAACTAT -1 797. 3300 L3 QSDYSYPYT Q95- T103 798. 3300 L3 CAGAGTGATTACTCATATCCTTACACA -1 799. 3300 L2 WAS W56-S58 800. 3300 L2 TGGGCTAGT -1 801. 3300 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R114- C220 802. 3300 CL AGGACTGTGCCGCTCCAAGTGTCTTCATTTTTCCACCCTCCGACGAACAGCTGAAGTCCGGCACAGCCTCTGT- GGTCTGTCTGCTGAACAATTTTTATCCACGCGAGGCC -1 AAGGTGCAGTGGAAAGTCGATAATGCTCTGCAGAGCGGGAACTCCCAGGAGTCTGTGACCGAACAGGACAG- TAAGGATTCAACATACAGCCTGTCAAGCACTCTGACC CTGTCTAAAGCAGATTATGAGAAGCACAAAGTGTACGCCTGCGAAGTCACTCATCAGGGACTGTCCTCTCC- CGTGACCAAGAGCTTCAACAGAGGCGAATGT 803. 2305 Full QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM -1 DYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ- S5GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP- REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFF- LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 804. 2305 Full CAGGTCCAGCTGCAGCAGAGCGGAGCCGAACTGGTCAGACCCGGCAGCTCCGTGAAAATCAGTTGCAAGGCTT- CAGGCTATGCATTCTCTAGTTACTGGATGAACTGGG -1 TGAAGCAGAGGCCTGGGCAGGGACTGGAATGGATCGGGCAGATTTGGCCAGGCGACGGGGATACTAACTAT- AATGGGAAGTTCAAAGGAAAGGCCACTCTGACCGCT GACGAGTCAAGCTCCACCGCCTATATGCAGCTGTCTAGTCTGGCATCTGAGGATAGTGCCGTGTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCCGCTACTATTA CGCTATGGACTATTGGGGACAGGGCACCACAGTCACTGTGTCAAGCGCTAGCACCAAAGGGCCTTCCGTGT- TTCCACTGGCACCCTCCTCTAAGAGCACTTCCGGAGGAA CCGCAGCTCTGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGTCACAGTGTCATGGAACAGCGGAGCA- CTGACCAGCGGAGTCCACACATTTCCTGCCGTGCTGCA GAGTTCAGGCCTGTATTCCCTGAGCTCCGTGGTCACAGTGCCATCTAGTTCACTGGGGACACAGACTTACA- TCTGCAACGTGAATCACAAACCATCCAATACTAAGGTCG ACAAGAAAGTGGAACCCAAATCTTGTGATAAGACCCATACATGCCCCCCTTGTCCTGCTCCAGAGCTGCTG- GGAGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAG GACACTCTGATGATTAGCCGAACACCAGAAGTCACTTGCGTGGTCGTGGACGTGAGCCACGAGGACCCCGA- AGTCAAGTTCAACTGGTACGTGGATGGGGTCGAGGTG CATAATGCCAAAACCAAGCCCAGGGAGGAACAGTATAATTCTACATACCGCGTCGTGAGTGTCCTGACTGT- GCTGCACCAGGACTGGCTGAACGGAAAGGAGTACAAAT GCAAGGTGTCCAACAAGGCACTGCCCGCCCCTATCGAGAAGACCATTTCTAAAGCAAAGGGCCAGCCTCGA- GAACCACAGGTCTATGTGCTGCCTCCAAGTCGGGACGA GCTGACAAAAAACCAGGTCAGCCTGCTGTGTCTGGTGAAGGGGTTCTACCCCTCCGATATTGCCGTGGAGT- GGGAATCTAATGGACAGCCTGAAAACAATTATCTGACCT GGCCCCCTGTGCTGGACTCCGATGGATCTTTCTTTCTGTACTCAAAACTGACAGTGGATAAGAGCAGGTGG- CAGCAGGGCAACGTCTTTTCTTGTAGTGTGATGCATGAG GCCCTGCACAATCATTACACCCAGAAATCACTGAGCCTGTCCCCCGGCAAG 805. 2305 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q1-S124 DYWGQGTTVTVSS 806. 2305 VH CAGGTCCAGCTGCAGCAGAGCGGAGCCGAACTGGTCAGACCCGGCAGCTCCGTGAAAATCAGTTGCAAGGCTT- CAGGCTATGCATTCTCTAGTTACTGGATGAACTGGG -1 TGAAGCAGAGGCCTGGGCAGGGACTGGAATGGATCGGGCAGATTTGGCCAGGCGACGGGGATACTAACTAT- AATGGGAAGTTCAAAGGAAAGGCCACTCTGACCGCT GACGAGTCAAGCTCCACCGCCTATATGCAGCTGTCTAGTCTGGCATCTGAGGATAGTGCCGTGTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCCGCTACTATTA CGCTATGGACTATTGGGGACAGGGCACCACAGTCACTGTGTCAAGC 807. 2305 H1 GYAFSSYW G26-W33 808. 2305 H1 GGCTATGCATTCTCTAGTTACTGG -1 809. 2305 H3 ARRETTTVGRYYYAMDY A97-Y113 810. 2305 H3 GCTCGGAGAGAAACCACAACTGTGGGCCGCTACTATTACGCTATGGACTAT -1 811. 2305 H2 IWPGDGDT I51-T58 812. 2305 H2 ATTTGGCCAGGCGACGGGGATACT -1

813. 2305 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A125- V222 814. 2305 CH1 GCTAGCACCAAAGGGCCTTCCGTGTTTCCACTGGCACCCTCCTCTAAGAGCACTTCCGGAGGAACCGCAGCTC- TGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGT CACAGTGTCATGGAACAGCGGAGCACTGACCAGCGGAGTCCACACATTTCCTGCCGTGCTGCAGAGTTCAG- GCCTGTATTCCCTGAGCTCCGTGGTCACAGTGCCATCTA GTTCACTGGGGACACAGACTTACATCTGCAACGTGAATCACAAACCATCCAATACTAAGGTCGACAAGAAA- GTG -1 815. 2305 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A238- K347 816. 2305 CH2 GCTCCAGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAGGACACTCTGATGATTAGCC- GAACACCAGAAGTCACTTGCGTGGTCGTGGACGTGA -1 GCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGATGGGGTCGAGGTGCATAATGCCAAAACCAAG- CCCAGGGAGGAACAGTATAATTCTACATACCGCGTCG TGAGTGTCCTGACTGTGCTGCACCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGTCCAACAAG- GCACTGCCCGCCCCTATCGAGAAGACCATTTCTAAAGC AAAG 817. 2305 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G348- G453 818. 2305 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGCTGCCTCCAAGTCGGGACGAGCTGACAAAAAACCAGGTCAGCC- TGCTGTGTCTGGTGAAGGGGTTCTACCCCTCCGATA -1 TTGCCGTGGAGTGGGAATCTAATGGACAGCCTGAAAACAATTATCTGACCTGGCCCCCTGTGCTGGACTCC- GATGGATCTTTCTTTCTGTACTCAAAACTGACAGTGGATA AGAGCAGGTGGCAGCAGGGCAACGTCTTTTCTTGTAGTGTGATGCATGAGGCCCTGCACAATCATTACACC- CAGAAATCACTGAGCCTGTCCCCCGGC 819. -2 Full DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTL- TISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIF -1 PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH- KVYACEVTHQGLSSPVTKSFNRGEC 820. -2 Full GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGG- CAAGTCAGGACGTTAACACCGCTGTAGCTTGGTATCA -1 GCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCTTTTTGTACAGTGGGGTCCCATCAA- GGTTCAGTGGCAGTCGATCTGGGACAGATTTCACTCTCAC CATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGCATTACACTACCCCACCCACTT- TCGGCCAAGGGACCAAAGTGGAGATCAAACGAACTGTGG CTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGC- CTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGT GGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAAGAGAGTGTCACAGAGCAGGACAGCAAGGACAGC- ACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAG CAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAG- AGCTTCAACAGGGGAGAGTGT 821. -2 VL DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFS- GSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK D1-K107 822. -2 VL GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATC- ACTTGCCGGGCAAGTCAGGACGTTAACACCGCTGTAGCTTGGTATCA -1 GCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCTTTTTGTACAGTGGGGTCCCATCAA- GGTTCAGTGGCAGTCGATCTGGGACAGATTTCACTCTCAC CATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAACAGCATTACACTACCCCACCCACTT- TCGGCCAAGGGACCAAAGTGGAGATCAAA 823. -2 L1 QDVNTA Q27-A32 824. -2 L1 CAGGACGTTAACACCGCT -1 825. -2 L3 QQHYTTPPT Q89-T97 826. -2 L3 CAACAGCATTACACTACCCCACCCACT -1 827. -2 L2 SAS S50-S52 828. -2 L2 TCTGCATCC -1 829. -2 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD- STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R108- C214 830. -2 CL CGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGA- ACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCC -1 AAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAAGAGAGTGTCACAGAGCAGGACAG- CAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACG CTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCC- CGTCACAAAGAGCTTCAACAGGGGAGAGTGT 831. 5238 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY CLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG 832. 5238 Full CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCTGAAGATGCCGCTACATACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTGCAGCTGCAGCAGTCCGGAGCTGAGCTGGCA- CGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGC CAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGGCAGGGACTGGAATGGA- TCGGGTACATTAATCCTAGCCGAGGATACACAAACTAC AACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAGATAAGAGCTCCTCTACCGCATATATGCAGCTGAG- TTCACTGACATCTGAGGACAGTGCCGTGTACTATTGCGC TAGGTACTATGACGATCACTACTGTCTGGATTATTGGGGCCAGGGGACTACCCTGACCGTGAGCTCCGCAG- CCGAACCTAAATCTAGTGACAAGACTCATACCTGCCCCC CTTGTCCAGCACCAGAGCTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTG- ATGATCTCCCGGACACCTGAAGTCACTTGCGTGGTCGTG GACGTGTCTCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCCACATAT CGCGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAG- CAACAAGGCCCTGCCTGCTCCAATCGAGAAGACAATTA GCAAAGCCAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAA- AACCAGGTCAGTCTGCTGTGTCTGGTGAAGGGCTTCTA TCCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTG- TCCTGGACTCAGATGGGAGCTTCTTTCTGTATAGTAAAC TGACCGTGGACAAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCAC- AATCATTACACCCAGAAGTCTCTGAGTCTGTCACCCGGC 833. 5238 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q1-N106 834. 5238 VL CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCTGAAGATGCCGCTACATACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAAT 835. 5238 L1 SSVSY S27-Y31 836. 5238 L1 TCCTCTGTGAGCTAC -1 837. 5238 L3 QQWSSNPFT Q88-T96 838. 5238 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 839. 5238 L2 DTS D49-S51 840. 5238 L2 GACACATCC -1 841. 5238 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYCLDYWGQ Q122- GTTLTVSS S240 842. 5238 VH CAGGTGCAGCTGCAGCAGTCCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGCCA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGGCAGGGACTGGAATGGATCGGGTACATTAATCCTAGCCGAGGATACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCTACTCTGACCACA GATAAGAGCTCCTCTACCGCATATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCCGTGTACTATTG- CGCTAGGTACTATGACGATCACTACTGTCTGGATTATTGG GGCCAGGGGACTACCCTGACCGTGAGCTCC 843. 5238 H1 GYTFTRYT G147- T154 844. 5238 H1 GGCTACACCTTCACACGGTATACC -1 845. 5238 H3 ARYYDDHYCLDY A218- Y229 846. 5238 H3 GCTAGGTACTATGACGATCACTACTGTCTGGATTAT -1 847. 5238 H2 INPSRGYT I172- T179 848. 5238 H2 ATTAATCCTAGCCGAGGATACACA -1 849. 5238 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 850. 5238 CH2 GCACCAGAGCTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGCGTGGTCGTGGACGTGTC TCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CCCTGCCTGCTCCAATCGAGAAGACAATTAGCAAAGCC AAG -1 851. 5238 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 852. 5238 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGCTTCTATCCAAGCGATA TTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTCCTGGACTCA- GATGGGAGCTTCTTTCTGTATAGTAAACTGACCGTGGAC AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAGTCTCTGAGTCTGTCACCCGGC -1 853. 2167 Full QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEINGGGGSGG -1 GGSGGGGSQVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQK- FKDKATLTTDK5SSTAYMQLSSLTSEDSAVYYCARYYDDHY SLDYWGQGTTLTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE- DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWES- NGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK 854. 2167 Full CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAATGGCGGAGGAG GCTCCGGAGGAGGAGGGTCTGGAGGAGGAGGAAGTCAGGTGCAGCTGCAGCAGAGCGGAGCTGAGCTGGCA- CGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGC CAGCGGCTACACCTTCACACGGTATACCATGCATTGGGTGAAACAGAGACCCGGGCAGGGACTGGAATGGA- TCGGGTACATTAATCCTTCCCGAGGATACACAAACTAC AACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAGATAAGAGCTCCTCTACCGCATATATGCAGCTGAG- TTCACTGACATCTGAGGACAGTGCCGTGTACTATTGCGC TAGGTACTATGACGATCACTACTCCCTGGATTATTGGGGCCAGGGGACTACCCTGACAGTGAGCTCCGCAG- CCGAACCTAAATCTAGTGACAAGACTCATACCTGCCCCC CTTGTCCAGCACCAGAGCTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTG- ATGATCTCCCGGACACCTGAAGTCACTTGTGTGGTCGTG GACGTGTCTCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAA- AACCAAGCCCAGGGAGGAACAGTACAACTCCACATATC GCGTCGTGTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGC- AACAAGGCCCTGCCTGCTCCAATCGAGAAGACAATTAG CAAAGCCAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAA- ACCAGGTCAGTCTGCTGTGTCTGGTGAAGGGCTTCTAT CCAAGCGATATTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGT- CCTGGACTCAGATGGGAGCTTCTTTCTGTATAGTAAACT GACCGTGGACAAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACA- ATCATTACACCCAGAAATCTCTGAGTCTGTCACCCGGCA AG

855. 2167 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q1-N106 856. 2167 VL CAGATCGTCCTGACACAGAGCCCAGCAATCATGTCAGCCAGCCCCGGCGAGAAAGTCACAATGACTTGCTCAG- CAAGCTCCTCTGTGAGCTACATGAACTGGTATCAGCA -1 GAAAAGCGGAACCTCCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCTTCTGGAGTGCCTGCACACT- TCAGGGGCAGCGGCTCTGGGACCAGTTATTCACTGACA ATTTCCGGCATGGAGGCTGAAGATGCCGCTACCTACTATTGCCAGCAGTGGAGTTCAAACCCATTCACTTT- TGGATCTGGCACCAAGCTGGAAATTAAT 857. 2167 L1 SSVSY S27-Y31 858. 2167 L1 TCCTCTGTGAGCTAC -1 859. 2167 L3 QQWSSNPFT Q88-T96 860. 2167 L3 CAGCAGTGGAGTTCAAACCCATTCACT -1 861. 2167 L2 DTS D49-S51 862. 2167 L2 GACACATCC -1 863. 2167 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ Q122- GTTLTVSS S240 864. 2167 VH CAGGTGCAGCTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCCTGTAAGGCCA- GCGGCTACACCTTCACACGGTATACCATGCATTGG -1 GTGAAACAGAGACCCGGGCAGGGACTGGAATGGATCGGGTACATTAATCCTTCCCGAGGATACACAAACTA- CAACCAGAAGTTTAAAGACAAGGCTACTCTGACCACAG ATAAGAGCTCCTCTACCGCATATATGCAGCTGAGTTCACTGACATCTGAGGACAGTGCCGTGTACTATTGC- GCTAGGTACTATGACGATCACTACTCCCTGGATTATTGGG GCCAGGGGACTACCCTGACAGTGAGCTCC 865. 2167 H1 GYTFTRYT G147- T154 866. 2167 H1 GGCTACACCTTCACACGGTATACC -1 867. 2167 H3 ARYYDDHYSLDY A218- Y229 868. 2167 H3 GCTAGGTACTATGACGATCACTACTCCCTGGATTAT -1 869. 2167 H2 INPSRGYT I172- T179 870. 2167 H2 ATTAATCCTTCCCGAGGATACACA -1 871. 2167 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A258- K367 872. 2167 CH2 GCACCAGAGCTGCTGGGAGGACCTAGCGTGTTCCTGTTTCCACCCAAACCAAAGGATACTCTGATGATCTCCC- GGACACCTGAAGTCACTTGTGTGGTCGTGGACGTGTC -1 TCACGAGGACCCCGAAGTCAAGTTTAACTGGTACGTGGACGGCGTCGAGGTGCATAATGCCAAAACCAAGC- CCAGGGAGGAACAGTACAACTCCACATATCGCGTCGT GTCTGTCCTGACTGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGAGCAACAAGG- CCCTGCCTGCTCCAATCGAGAAGACAATTAGCAAAGCC AAG 873. 2167 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G368- G473 874. 2167 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGCTGCCTCCATCTCGGGACGAGCTGACTAAAAACCAGGTCAGTC- TGCTGTGTCTGGTGAAGGGCTTCTATCCAAGCGATA -1 TTGCTGTGGAGTGGGAATCCAATGGGCAGCCCGAAAACAATTACCTGACTTGGCCCCCTGTCCTGGACTCA- GATGGGAGCTTCTTTCTGTATAGTAAACTGACCGTGGAC AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGCTGTTCCGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAATCTCTGAGTCTGTCACCCGGC 875. 3320 Full EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTIS- RDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYW -1 AYWGQGTLVTVSSGGGGSGGGGSGGGGSQTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPG- QAPRGLIGGTKFLAPGTPARFSGSLLGGKAALTLSGVQPE DEAEYYCVLWYSNRWVFGGGTKLTVLAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPE- VTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGF- YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSR WQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 876. 3320 Full GAAGTCCAGCTGGTCGAGTCCGGAGGAGGACTGGTGCAGCCAGGAGGGTCACTGAAACTGAGCTGCGCCGCTT- CCGGCTTCACTTTTAACAAGTATGCAATGAATTGG -1 GTGCGGCAGGCACCAGGGAAGGGACTGGAATGGGTGGCCCGGATCAGATCTAAGTACAACAACTACGCTAC- CTACTATGCAGACAGTGTGAAGGATAGGTTCACAATT TCTCGCGACGATAGTAAAAACACTGCTTACCTGCAGATGAACAATCTGAAGACAGAGGACACTGCAGTCTA- CTATTGCGTGAGACACGGAAACTTTGGCAATAGCTACAT CTCCTATTGGGCATACTGGGGACAGGGAACCCTGGTCACAGTGAGCTCCGGAGGAGGAGGCAGCGGAGGAG- GAGGCTCTGGGGGAGGCGGGAGTCAGACTGTGGTC ACCCAGGAGCCCTCACTGACAGTCAGCCCTGGAGGCACTGTGACCCTGACATGTGGGTCTAGTACCGGAGC- CGTGACATCTGGCAACTATCCCAATTGGGTGCAGCAGA AACCTGGACAGGCTCCACGAGGACTGATTGGAGGAACAAAGTTCCTGGCCCCCGGAACTCCTGCTCGATTT- TCCGGCTCTCTGCTGGGAGGGAAAGCAGCACTGACCCT GAGCGGAGTGCAGCCTGAGGATGAAGCCGAGTACTATTGCGTGCTGTGGTACAGCAACAGATGGGTGTTCG- GAGGCGGGACAAAGCTGACTGTGCTGGCTGCAGAGC CAAAGTCAAGCGACAAAACTCACACCTGCCCACCTTGTCCAGCTCCAGAAGCAGCTGGAGGACCATCCGTG- TTCCTGTTTCCACCCAAGCCCAAAGATACACTGATGATCT CTCGCACTCCCGAGGTCACCTGTGTGGTCGTGAGTGTGTCACACGAAGACCCTGAGGTCAAGTTTAACTGG- TACGTGGATGGCGTCGAAGTGCATAATGCCAAGACCAA ACCTCGAGAGGAACAGTATAATTCAACTTACCGGGTCGTGAGCGTCCTGACCGTGCTGCATCAGGACTGGC- TGAACGGAAAGGAGTACAAGTGCAAAGTGAGCAATAA GGCACTGCCTGCCCCAATCGAAAAAACCATTAGCAAGGCTAAAGGGCAGCCAAGAGAGCCCCAGGTCTACG- TGTATCCTCCAAGCAGGGACGAACTGACCAAGAACCA GGTCTCCCTGACATGTCTGGTGAAAGGGTTCTATCCTAGTGATATTGCAGTGGAATGGGAGTCAAATGGAC- AGCCAGAGAACAATTACAAGACCACACCCCCTGTGCTG GACTCTGATGGCAGTTTCGCACTGGTCTCCAAGCTGACCGTGGATAAATCTAGGTGGCAGCAGGGGAACGT- CTTTAGCTGTTCCGTGATGCATGAAGCCCTGCACAATCA TTACACACAGAAGTCTCTGAGTCTGTCACCCGGCAAA 877. 3320 VH EVQLVESGGGLVQPGGSLKLSCAASGFTFNKYAMNWVRQAPGKGLEWVARIRSKYNNYATYYADSVKDRFTIS- RDDSKNTAYLQMNNLKTEDTAVYYCVRHGNFGNSYISYW E1-S125 AYWGQGTLVTVSS 878. 3320 VH GAAGTCCAGCTGGTCGAGTCCGGAGGAGGACTGGTGCAGCCAGGAGGGTCACTGAAACTGAGCTGCGCCGCTT- CCGGCTTCACTTTTAACAAGTATGCAATGAATTGG -1 GTGCGGCAGGCACCAGGGAAGGGACTGGAATGGGTGGCCCGGATCAGATCTAAGTACAACAACTACGCTAC- CTACTATGCAGACAGTGTGAAGGATAGGTTCACAATT TCTCGCGACGATAGTAAAAACACTGCTTACCTGCAGATGAACAATCTGAAGACAGAGGACACTGCAGTCTA- CTATTGCGTGAGACACGGAAACTTTGGCAATAGCTACAT CTCCTATTGGGCATACTGGGGACAGGGAACCCTGGTCACAGTGAGCTCC 879. 3320 H1 GFTFNKYA G26-A33 880. 3320 H1 GGCTTCACTTTTAACAAGTATGCA -1 881. 3320 H3 VRHGNFGNSYISYWAY V99-Y114 882. 3320 H3 GTGAGACACGGAAACTTTGGCAATAGCTACATCTCCTATTGGGCATAC -1 883. 3320 H2 IRSKYNNYAT I51-T60 884. 3320 H2 ATCAGATCTAAGTACAACAACTACGCTACC -1 885. 3320 VL QTVVTQEPSLTVSPGGTVTLTCGSSTGAVTSGNYPNWVQQKPGQAPRGLIGGTKFLAPGTPARFSGSLLGGKA- ALTLSGVQPEDEAEYYCVLWYSNRWVFGGGTKLTVL Q141- L249 886. 3320 VL CAGACTGTGGTCACCCAGGAGCCCTCACTGACAGTCAGCCCTGGAGGCACTGTGACCCTGACATGTGGGTCTA- GTACCGGAGCCGTGACATCTGGCAACTATCCCAATTG -1 GGTGCAGCAGAAACCTGGACAGGCTCCACGAGGACTGATTGGAGGAACAAAGTTCCTGGCCCCCGGAACTC- CTGCTCGATTTTCCGGCTCTCTGCTGGGAGGGAAAGCA GCACTGACCCTGAGCGGAGTGCAGCCTGAGGATGAAGCCGAGTACTATTGCGTGCTGTGGTACAGCAACAG- ATGGGTGTTCGGAGGCGGGACAAAGCTGACTGTGCTG 887. 3320 L1 TGAVTSGNY T166- Y174 888. 3320 L1 ACCGGAGCCGTGACATCTGGCAACTAT -1 889. 3320 L3 VLWYSNRWV V231- V239 890. 3320 L3 GTGCTGTGGTACAGCAACAGATGGGTG -1 891. 3320 L2 GTK G192- K194 892. 3320 L2 GGAACAAAG -1 893. 3320 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A267- K376 894. 3320 CH2 GCTCCAGAAGCAGCTGGAGGACCATCCGTGTTCCTGTTTCCACCCAAGCCCAAAGATACACTGATGATCTCTC- GCACTCCCGAGGTCACCTGTGTGGTCGTGAGTGTGTC -1 ACACGAAGACCCTGAGGTCAAGTTTAACTGGTACGTGGATGGCGTCGAAGTGCATAATGCCAAGACCAAAC- CTCGAGAGGAACAGTATAATTCAACTTACCGGGTCGTG AGCGTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAGTGCAAAGTGAGCAATAAGGC- ACTGCCTGCCCCAATCGAAAAAACCATTAGCAAGGCT AAA 895. 3320 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G377- G482 896. 3320 CH3 GGGCAGCCAAGAGAGCCCCAGGTCTACGTGTATCCTCCAAGCAGGGACGAACTGACCAAGAACCAGGTCTCCC- TGACATGTCTGGTGAAAGGGTTCTATCCTAGTGATA -1 TTGCAGTGGAATGGGAGTCAAATGGACAGCCAGAGAACAATTACAAGACCACACCCCCTGTGCTGGACTCT- GATGGCAGTTTCGCACTGGTCTCCAAGCTGACCGTGGA TAAATCTAGGTGGCAGCAGGGGAACGTCTTTAGCTGTTCCGTGATGCATGAAGCCCTGCACAATCATTACA- CACAGAAGTCTCTGAGTCTGTCACCCGGC 897. 5241 Full QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM -1 DYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ- SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP- REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFA- LVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 898. 5241 Full CAGGTCCAGCTGCAGCAGAGCGGAGCCGAACTGGTCAGACCCGGCAGCTCCGTGAAAATCAGTTGCAAGGCTT- CAGGCTATGCATTCTCTAGTTACTGGATGAACTGGG -1 TGAAGCAGAGGCCTGGGCAGGGACTGGAATGGATCGGGCAGATTTGGCCAGGCGACGGGGATACAAACTAT- AATGGGAAGTTCAAAGGAAAGGCCACACTGACTGCT GACGAGTCAAGCTCCACTGCATATATGCAGCTGTCTAGTCTGGCATCTGAGGATAGTGCCGTGTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCCGCTACTATTA CGCCATGGACTATTGGGGACAGGGCACCACAGTCACAGTGTCAAGCGCTAGCACTAAAGGGCCTTCCGTGT- TTCCACTGGCACCCTCCTCTAAGAGCACATCCGGAGGA ACTGCAGCTCTGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGTCACAGTGTCATGGAACAGCGGCGC- ACTGACTAGCGGGGTCCACACCTTTCCTGCCGTGCTGCA GAGTTCAGGCCTGTATTCCCTGAGCTCCGTGGTCACCGTGCCATCTAGTTCACTGGGGACCCAGACATACA- TCTGCAACGTGAATCACAAACCATCCAATACAAAGGTCG ACAAGAAAGTGGAACCCAAATCTTGTGATAAGACTCATACCTGCCCCCCTTGTCCTGCTCCAGAGCTGCTG- GGAGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAG GACACACTGATGATTAGCCGAACCCCAGAAGTGACATGCGTGGTCGTGGACGTGAGCCACGAGGACCCCGA- AGTCAAATTCAACTGGTACGTGGATGGGGTCGAGGTG CATAATGCCAAAACCAAGCCCAGGGAGGAACAGTATAATTCTACTTACCGCGTCGTGAGTGTCCTGACCGT- GCTGCACCAGGACTGGCTGAACGGAAAGGAGTACAAAT GCAAGGTGTCCAACAAGGCACTGCCCGCCCCTATCGAGAAGACCATTTCTAAAGCTAAGGGCCAGCCTCGA- GAACCACAGGTCTATGTGTACCCTCCAAGTCGGGACGA GCTGACCAAAAACCAGGTCAGCCTGACATGTCTGGTGAAGGGGTTTTATCCCTCCGATATTGCAGTGGAGT- GGGAATCTAATGGACAGCCTGAAAACAATTACAAGACT ACCCCCCCTGTGCTGGACTCCGATGGATCTTTCGCCCTGGTCTCAAAACTGACTGTGGATAAGAGCAGGTG- GCAGCAGGGCAACGTCTTTTCTTGTAGTGTGATGCATGA GGCTCTGCACAATCATTACACCCAGAAGTCACTGAGCCTGTCCCCCGGC 899. 5241 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q1-S124 DYWGQGTTVTVSS 900. 5241 VH CAGGTCCAGCTGCAGCAGAGCGGAGCCGAACTGGTCAGACCCGGCAGCTCCGTGAAAATCAGTTGCAAGGCTT- CAGGCTATGCATTCTCTAGTTACTGGATGAACTGGG -1 TGAAGCAGAGGCCTGGGCAGGGACTGGAATGGATCGGGCAGATTTGGCCAGGCGACGGGGATACAAACTAT- AATGGGAAGTTCAAAGGAAAGGCCACACTGACTGCT GACGAGTCAAGCTCCACTGCATATATGCAGCTGTCTAGTCTGGCATCTGAGGATAGTGCCGTGTACTTTTG- CGCTCGGAGAGAAACCACAACTGTGGGCCGCTACTATTA

CGCCATGGACTATTGGGGACAGGGCACCACAGTCACAGTGTCAAGC 901. 5241 H1 GYAFSSYW G26-W33 902. 5241 H1 GGCTATGCATTCTCTAGTTACTGG -1 903. 5241 H3 ARRETTTVGRYYYAMDY A97-Y113 904. 5241 H3 GCTCGGAGAGAAACCACAACTGTGGGCCGCTACTATTACGCCATGGACTAT -1 905. 5241 H2 IWPGDGDT I51-T58 906. 5241 H2 ATTTGGCCAGGCGACGGGGATACA -1 907. 5241 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A125- V222 908. 5241 CH1 GCTAGCACTAAAGGGCCTTCCGTGTTTCCACTGGCACCCTCCTCTAAGAGCACATCCGGAGGAACTGCAGCTC- TGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGT -1 CACAGTGTCATGGAACAGCGGCGCACTGACTAGCGGGGTCCACACCTTTCCTGCCGTGCTGCAGAGTTCAG- GCCTGTATTCCCTGAGCTCCGTGGTCACCGTGCCATCTA GTTCACTGGGGACCCAGACATACATCTGCAACGTGAATCACAAACCATCCAATACAAAGGTCGACAAGAAA- GTG 909. 5241 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A238- K347 910. 5241 CH2 GCTCCAGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAGGACACACTGATGATTAGCC- GAACCCCAGAAGTGACATGCGTGGTCGTGGACGTGA -1 GCCACGAGGACCCCGAAGTCAAATTCAACTGGTACGTGGATGGGGTCGAGGTGCATAATGCCAAAACCAAG- CCCAGGGAGGAACAGTATAATTCTACTTACCGCGTCGT GAGTGTCCTGACCGTGCTGCACCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGTCCAACAAGG- CACTGCCCGCCCCTATCGAGAAGACCATTTCTAAAGCT AAG 911. 5241 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G348- G453 912. 5241 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGTACCCTCCAAGTCGGGACGAGCTGACCAAAAACCAGGTCAGCC- TGACATGTCTGGTGAAGGGGTTTTATCCCTCCGATAT -1 TGCAGTGGAGTGGGAATCTAATGGACAGCCTGAAAACAATTACAAGACTACCCCCCCTGTGCTGGACTCCG- ATGGATCTTTCGCCCTGGTCTCAAAACTGACTGTGGATA AGAGCAGGTGGCAGCAGGGCAACGTCTTTTCTTGTAGTGTGATGCATGAGGCTCTGCACAATCATTACACC- CAGAAGTCACTGAGCCTGTCCCCCGGC 913. 5242 Full QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM -1 DYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ- SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKP- REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFA- LVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 914. 5242 Full CAGGTCCAGCTGCAGCAGTCCGGAGCCGAACTGGTCAGACCCGGCAGCTCCGTGAAAATCAGCTGCAAGGCCT- CCGGCTATGCTTTCTCTAGTTACTGGATGAACTGGG -1 TGAAGCAGAGGCCTGGGCAGGGACTGGAATGGATCGGGCAGATTTGGCCAGGCGACGGGGATACAAACTAT- AATGGGAAGTTCAAAGGAAAGGCAACACTGACTGCC GACGAGTCAAGCTCCACTGCTTATATGCAGCTGTCTAGTCTGGCTTCAGAGGATAGCGCAGTGTACTTTTG- CGCCCGGAGAGAAACCACAACTGTGGGCCGCTACTATTA CGCAATGGACTATTGGGGACAGGGCACCACAGTCACAGTGTCAAGCGCCTCTACTAAAGGGCCTAGTGTGT- TTCCACTGGCTCCCTCCTCTAAGAGCACATCCGGAGGA ACTGCAGCTCTGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGTCACAGTGTCCTGGAACTCTGGCGC- TCTGACTAGCGGGGTCCACACCTTTCCTGCAGTGCTGCA GAGTTCAGGCCTGTATAGCCTGAGCTCCGTGGTCACCGTGCCATCTAGTTCACTGGGGACCCAGACATACA- TCTGCAACGTGAATCACAAACCAAGCAATACAAAGGTCG ACAAGAAAGTGGAACCCAAAAGCTGTGATAAGACTCATACCTGCCCCCCTTGTCCTGCACCAGAGGCAGCA- GGAGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAG GACACACTGATGATTTCCCGAACCCCAGAAGTGACATGCGTGGTCGTGTCTGTGAGTCACGAGGACCCCGA- AGTCAAATTCAACTGGTACGTGGATGGGGTCGAGGTGC ATAATGCCAAAACCAAGCCCAGGGAGGAACAGTATAATTCAACTTACCGCGTCGTGAGCGTCCTGACCGTG- CTGCACCAGGATTGGCTGAACGGAAAGGAGTACAAAT GCAAGGTGTCCAACAAGGCTCTGCCCGCACCTATCGAGAAGACCATTTCTAAAGCTAAGGGCCAGCCTCGA- GAACCACAGGTCTATGTGTACCCTCCATCCCGGGACGA GCTGACCAAAAACCAGGTCTCTCTGACATGTCTGGTGAAGGGGTTTTATCCCAGTGATATTGCCGTGGAGT- GGGAAAGCAATGGACAGCCTGAAAACAATTACAAGACT ACCCCCCCTGTGCTGGACAGTGATGGATCATTCGCACTGGTCTCCAAACTGACTGTGGACAAGTCTAGGTG- GCAGCAGGGCAACGTCTTTTCATGTAGCGTGATGCATGA GGCCCTGCACAATCATTACACCCAGAAGTCCCTGTCTCTGAGTCCCGGC 915. 5242 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q1-S124 DYWGQGTTVTVSS 916. 5242 VH CAGGTCCAGCTGCAGCAGTCCGGAGCCGAACTGGTCAGACCCGGCAGCTCCGTGAAAATCAGCTGCAAGGCCT- CCGGCTATGCTTTCTCTAGTTACTGGATGAACTGGG -1 TGAAGCAGAGGCCTGGGCAGGGACTGGAATGGATCGGGCAGATTTGGCCAGGCGACGGGGATACAAACTAT- AATGGGAAGTTCAAAGGAAAGGCAACACTGACTGCC GACGAGTCAAGCTCCACTGCTTATATGCAGCTGTCTAGTCTGGCTTCAGAGGATAGCGCAGTGTACTTTTG- CGCCCGGAGAGAAACCACAACTGTGGGCCGCTACTATTA CGCAATGGACTATTGGGGACAGGGCACCACAGTCACAGTGTCAAGC 917. 5242 H1 GYAFSSYW G26-W33 918. 5242 H1 GGCTATGCTTTCTCTAGTTACTGG -1 919. 5242 H3 ARRETTTVGRYYYAMDY A97-Y113 920. 5242 H3 GCCCGGAGAGAAACCACAACTGTGGGCCGCTACTATTACGCAATGGACTAT -1 921. 5242 H2 IWPGDGDT I51-T58 922. 5242 H2 ATTTGGCCAGGCGACGGGGATACA -1 923. 5242 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A125- V222 924. 5242 CH1 GCCTCTACTAAAGGGCCTAGTGTGTTTCCACTGGCTCCCTCCTCTAAGAGCACATCCGGAGGAACTGCAGCTC- TGGGATGTCTGGTGAAGGATTACTTCCCAGAGCCCGT -1 CACAGTGTCCTGGAACTCTGGCGCTCTGACTAGCGGGGTCCACACCTTTCCTGCAGTGCTGCAGAGTTCAG- GCCTGTATAGCCTGAGCTCCGTGGTCACCGTGCCATCTA GTTCACTGGGGACCCAGACATACATCTGCAACGTGAATCACAAACCAAGCAATACAAAGGTCGACAAGAAA- GTG 925. 5242 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A238- K347 926. 5242 CH2 GCACCAGAGGCAGCAGGAGGACCAAGCGTGTTCCTGTTTCCACCCAAACCTAAGGACACACTGATGATTTCCC- GAACCCCAGAAGTGACATGCGTGGTCGTGTCTGTGA -1 GTCACGAGGACCCCGAAGTCAAATTCAACTGGTACGTGGATGGGGTCGAGGTGCATAATGCCAAAACCAAG- CCCAGGGAGGAACAGTATAATTCAACTTACCGCGTCGT GAGCGTCCTGACCGTGCTGCACCAGGATTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGTCCAACAAGG- CTCTGCCCGCACCTATCGAGAAGACCATTTCTAAAGCT AAG 927. 5242 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G348- G453 928. 5242 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGTACCCTCCATCCCGGGACGAGCTGACCAAAAACCAGGTCTCTC- TGACATGTCTGGTGAAGGGGTTTTATCCCAGTGATAT -1 TGCCGTGGAGTGGGAAAGCAATGGACAGCCTGAAAACAATTACAAGACTACCCCCCCTGTGCTGGACAGTG- ATGGATCATTCGCACTGGTCTCCAAACTGACTGTGGAC AAGTCTAGGTGGCAGCAGGGCAACGTCTTTTCATGTAGCGTGATGCATGAGGCCCTGCACAATCATTACAC- CCAGAAGTCCCTGTCTCTGAGTCCCGGC 929. 5243 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGCGTKLEIKGGGGS -1 GGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQCLEWIGQIWPGDGDTNYN- GKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRET TTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC- VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYP- SDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPG 930. 5243 Full GATATTCAGCTGACTCAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACCATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGAGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGCCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACATACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGATGTGGCACTAAACTGGAAAT CAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGCG- GAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGCAAGGCATCTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGC- CAGTGTCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACAAACTATAATGGAAAGTTCAAAGGCAAGGCTACACTGACTGCAGACGAGTCAAGCTCCACTG- CTTATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCC GCTGTGTACTTTTGCGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAA CCCAAATCCTCTGATAAGACCCACACATGCCCTCCATGTCCAGCACCTGAGCTGCTGGGAGGACCAAGCGT- GTTCCTGTTTCCACCTAAACCTAAGGACACTCTGATGATC TCTCGGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTG- GTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAA AGCCTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTGTCTGTCCTGACCGTGCTGCATCAGGACTGG- CTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACA AGGCCCTGCCAGCTCCCATCGAGAAGACCATTTCCAAAGCTAAGGGCCAGCCTCGAGAACCACAGGTCTAT- GTGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCA GGTCTCCCTGACATGTCTGGTGAAGGGGTTTTATCCTTCTGATATTGCCGTGGAGTGGGAAAGTAATGGAC- AGCCAGAAAACAATTACAAAACTACCCCTCCAGTGCTGG ATTCTGACGGCAGTTTCGCACTGGTCAGTAAACTGACCGTGGATAAGTCACGGTGGCAGCAGGGGAACGTC- TTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCA TTACACACAGAAGAGCCTGTCCCTGTCTCCCGGC 931. 5243 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGCGTKLEIK D1-K111 932. 5243 VL GATATTCAGCTGACTCAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACCATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGAGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGCCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGGACT GATTTTACCCTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACATACCATTGCCAGCAGTCTACCGA- GGACCCCTGGACATTCGGATGTGGCACTAAACTGGAAAT CAAG 933. 5243 L1 QSVDYDGDSY Q27-Y36 934. 5243 L1 CAGTCAGTGGACTATGATGGAGACTCCTAT -1 935. 5243 L3 QQSTEDPWT Q93- T101 936. 5243 L3 CAGCAGTCTACCGAGGACCCCTGGACA -1 937. 5243 L2 DAS D54-S56 938. 5243 L2 GACGCCTCA -1 939. 5243 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQCLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 940. 5243 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGCAAGGCAT- CTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGCCAGTGTCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACAAACTAT- AATGGAAAGTTCAAAGGCAAGGCTACACTGACTGCA GACGAGTCAAGCTCCACTGCTTATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCCGCTGTGTACTTTTG- CGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCAATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 941. 5243 H1 GYAFSSYW G152- W159 942. 5243 H1 GGCTATGCCTTTTCTAGTTACTGG -1 943. 5243 H3 ARRETTTVGRYYYAMDY A223- Y239 944. 5243 H3 GCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCAATGGACTAC -1 945. 5243 H2 IWPGDGDT I177- T184 946. 5243 H2 ATTTGGCCCGGGGATGGAGACACA -1

947. 5243 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 948. 5243 CH2 GCACCTGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCTAAACCTAAGGACACTCTGATGATCTCTC- GGACACCCGAAGTCACTTGTGTGGTCGTGGATGTGAG -1 CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAAGC- CTAGGGAGGAACAGTATAACTCCACTTACCGCGTCGTG TCTGTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- CCTGCCAGCTCCCATCGAGAAGACCATTTCCAAAGCTA AG 949. 5243 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 950. 5243 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGTACCCACCCAGCCGGGACGAGCTGACCAAAAACCAGGTCTCCC- TGACATGTCTGGTGAAGGGGTTTTATCCTTCTGATAT -1 TGCCGTGGAGTGGGAAAGTAATGGACAGCCAGAAAACAATTACAAAACTACCCCTCCAGTGCTGGATTCTG- ACGGCAGTTTCGCACTGGTCAGTAAACTGACCGTGGAT AAGTCACGGTGGCAGCAGGGGAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACAC- ACAGAAGAGCCTGTCCCTGTCTCCCGGC 951. 2174 Full QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- K5SSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ -1 GTTLTV555STGGGGSGGGGSGGGGSDIQIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSP- KRWIYDTSKLASGVPAHFRGSGSGTSYSLTISGMEAEDAATYY CQQWSSNPFTFGSGTKLEINRAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVV- VDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIA- VEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK 952. 2174 Full CAGGTCCAGCTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCATGCAAGGCCA- GCGGCTACACCTTCACACGGTATACTATGCACTGG -1 GTGAAACAGAGACCCGGACAGGGCCTGGAATGGATCGGGTACATTAACCCTAGCCGAGGATACACCAACTA- CAACCAGAAGTTTAAAGACAAGGCTACCCTGACCACA GATAAGAGCTCCTCTACAGCATATATGCAGCTGAGTTCACTGACTTCTGAGGACAGTGCTGTGTACTATTG- TGCACGGTACTATGACGATCATTACTCCCTGGATTATTGG GGGCAGGGAACTACCCTGACCGTGAGCTCCTCTAGTACAGGAGGAGGAGGCAGTGGAGGAGGAGGGTCAGG- CGGAGGAGGAAGCGACATCCAGATTGTGCTGACAC AGTCTCCAGCAATCATGTCCGCCTCTCCCGGCGAGAAAGTCACTATGACCTGCTCCGCCTCAAGCTCCGTG- TCTTACATGAATTGGTATCAGCAGAAATCAGGAACCAGCC CCAAGAGATGGATCTACGACACATCCAAGCTGGCCTCTGGCGTGCCTGCTCACTTCAGGGGCAGTGGGTCA- GGAACTAGCTATTCCCTGACCATTAGCGGCATGGAGGC CGAAGATGCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAACCCATTCACATTTGGCAGCGGGACTAAGC- TGGAGATCAATAGGGCAGCCGAACCCAAATCAAGCGAC AAGACACATACTTGCCCCCCTTGTCCAGCACCAGAACTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCACC- CAAACCAAAGGATACACTGATGATTAGCCGCACCCCTGAG GTCACATGCGTGGTCGTGGACGTGAGCCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGT- CGAAGTGCATAATGCCAAAACCAAGCCTAGGGAGGAA CAGTACAACAGTACATATAGAGTCGTGTCAGTGCTGACCGTCCTGCACCAGGATTGGCTGAACGGCAAGGA- GTACAAATGCAAGGTGTCCAACAAGGCCCTGCCTGCTC CAATCGAGAAGACCATTTCTAAAGCAAAGGGGCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCC- CGGGACGAGCTGACTAAAAACCAGGTCTCTCTGACCTGT CTGGTGAAGGGCTTTTACCCATCTGATATTGCTGTCGAGTGGGAAAGTAATGGGCAGCCCGAGAACAATTA- TAAGACAACTCCCCCTGTGCTGGACTCCGATGGGTCTTT CGCCCTGGTCAGCAAACTGACAGTGGATAAGTCCAGATGGCAGCAGGGAAACGTCTTTTCTTGTAGTGTGA- TGCATGAAGCTCTGCACAATCATTACACTCAGAAATCAC TGAGCCTGTCCCCCGGCAAG 953. 2174 VH QVQLQQSGAELARPGASVKMSCKASGYTFTRYTMHWVKQRPGQGLEWIGYINPSRGYTNYNQKFKDKATLTTD- KSSSTAYMQLSSLTSEDSAVYYCARYYDDHYSLDYWGQ Q1-S119 GTTLTVSS 954. 2174 VH CAGGTCCAGCTGCAGCAGAGCGGAGCTGAGCTGGCACGACCAGGAGCAAGTGTGAAAATGTCATGCAAGGCCA- GCGGCTACACCTTCACACGGTATACTATGCACTGG -1 GTGAAACAGAGACCCGGACAGGGCCTGGAATGGATCGGGTACATTAACCCTAGCCGAGGATACACCAACTA- CAACCAGAAGTTTAAAGACAAGGCTACCCTGACCACA GATAAGAGCTCCTCTACAGCATATATGCAGCTGAGTTCACTGACTTCTGAGGACAGTGCTGTGTACTATTG- TGCACGGTACTATGACGATCATTACTCCCTGGATTATTGG GGGCAGGGAACTACCCTGACCGTGAGCTCC 955. 2174 H1 GYTFTRYT G26-T33 956. 2174 H1 GGCTACACCTTCACACGGTATACT -1 957. 2174 H3 ARYYDDHYSLDY A97-Y108 958. 2174 H3 GCACGGTACTATGACGATCATTACTCCCTGGATTAT -1 959. 2174 H2 INPSRGYT I51-T58 960. 2174 H2 ATTAACCCTAGCCGAGGATACACC -1 961. 2174 VL QIVLTQSPAIMSASPGEKVTMTCSASSSVSYMNWYQQKSGTSPKRWIYDTSKLASGVPAHFRGSGSGTSYSLT- ISGMEAEDAATYYCQQWSSNPFTFGSGTKLEIN Q140- N245 962. 2174 VL CAGATTGTGCTGACACAGTCTCCAGCAATCATGTCCGCCTCTCCCGGCGAGAAAGTCACTATGACCTGCTCCG- CCTCAAGCTCCGTGTCTTACATGAATTGGTATCAGCAG -1 AAATCAGGAACCAGCCCCAAGAGATGGATCTACGACACATCCAAGCTGGCCTCTGGCGTGCCTGCTCACTT- CAGGGGCAGTGGGTCAGGAACTAGCTATTCCCTGACCA TTAGCGGCATGGAGGCCGAAGATGCCGCTACCTACTATTGTCAGCAGTGGTCTAGTAACCCATTCACATTT- GGCAGCGGGACTAAGCTGGAGATCAAT 963. 2174 L1 SSVSY S166- Y170 964. 2174 L1 AGCTCCGTGTCTTAC -1 965. 2174 L3 QQWSSNPFT Q227- T235 966. 2174 L3 CAGCAGTGGTCTAGTAACCCATTCACA -1 967. 2174 L2 DTS D188- S190 968. 2174 L2 GACACATCC -1 969. 2174 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A264- K373 970. 2174 CH2 GCACCAGAACTGCTGGGAGGACCTTCCGTGTTCCTGTTTCCACCCAAACCAAAGGATACACTGATGATTAGCC- GCACCCCTGAGGTCACATGCGTGGTCGTGGACGTGAG -1 CCACGAGGACCCCGAAGTCAAGTTCAACTGGTACGTGGACGGCGTCGAAGTGCATAATGCCAAAACCAAGC- CTAGGGAGGAACAGTACAACAGTACATATAGAGTCGT GTCAGTGCTGACCGTCCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAATGCAAGGTGTCCAACAAGG- CCCTGCCTGCTCCAATCGAGAAGACCATTTCTAAAGCA AAG 971. 2174 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G374- G479 972. 2174 CH3 GGGCAGCCCCGAGAACCTCAGGTCTACGTGTATCCTCCATCCCGGGACGAGCTGACTAAAAACCAGGTCTCTC- TGACCTGTCTGGTGAAGGGCTTTTACCCATCTGATAT -1 TGCTGTCGAGTGGGAAAGTAATGGGCAGCCCGAGAACAATTATAAGACAACTCCCCCTGTGCTGGACTCCG- ATGGGTCTTTCGCCCTGGTCAGCAAACTGACAGTGGAT AAGTCCAGATGGCAGCAGGGAAACGTCTTTTCTTGTAGTGTGATGCATGAAGCTCTGCACAATCATTACAC- TCAGAAATCACTGAGCCTGTCCCCCGGC 973. 2175 Full DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGG -1 SGGGGSGGGGSQVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNY- NGKFKGKATLTADESSSTAYMQLSSLASEDSAVYFCARRE TTTVGRYYYAMDYWGQGTTVTVSSAAEPKSSDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT- CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFY- PSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 974. 2175 Full GACATTCAGCTGACCCAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACAATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGAACC GATTTTACACTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACAGA- GGACCCCTGGACTTTCGGCGGGGGAACCAAACTGGAAA TCAAGGGAGGAGGAGGCAGTGGCGGAGGAGGGTCAGGAGGAGGAGGAAGCCAGGTGCAGCTGCAGCAGAGC- GGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGA AAATTTCCTGTAAGGCATCTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGGTGAAGCAGAGGCCAGGA- CAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGA TGGAGACACAAACTATAATGGAAAGTTCAAAGGCAAGGCTACTCTGACCGCAGACGAGTCAAGCTCCACTG- CATATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCC GCTGTCTACTTTTGCGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCCATGGACTACTGGGG- CCAGGGGACCACAGTCACCGTGTCAAGCGCAGCCGAAC CCAAATCCTCTGATAAGACACACACTTGCCCTCCATGTCCAGCTCCTGAGCTGCTGGGAGGACCAAGCGTG- TTCCTGTTTCCACCTAAACCTAAGGACACTCTGATGATCT CTCGGACTCCCGAAGTCACCTGTGTGGTCGTGGATGTGAGCCACGAGGACCCTGAAGTCAAATTCAACTGG- TACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAA GCCTAGGGAGGAACAGTATAACTCCACATACCGCGTCGTGTCTGTCCTGACTGTGCTGCATCAGGACTGGC- TGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAA GGCCCTGCCAGCTCCCATCGAGAAGACCATTTCCAAAGCTAAGGGCCAGCCTCGAGAACCACAGGTCTATG- TGCTGCCACCCAGCCGGGACGAGCTGACAAAAAACCAG GTCTCCCTGCTGTGTCTGGTGAAGGGATTCTACCCTTCTGATATTGCAGTGGAGTGGGAAAGTAATGGCCA- GCCAGAAAACAATTATCTGACTTGGCCTCCAGTGCTGGA TTCTGACGGGAGTTTCTTTCTGTACAGTAAACTGACCGTGGATAAGTCACGGTGGCAGCAGGGAAACGTCT- TTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATT ACACCCAGAAAAGCCTGTCCCTGTCTCCCGGCAAG 975. 2175 VL DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRFSGSGSGT- DFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIK D1-K111 976. 2175 VL GACATTCAGCTGACCCAGAGTCCTGCTTCACTGGCAGTGAGCCTGGGACAGCGAGCAACAATCTCCTGCAAAG- CTAGTCAGTCAGTGGACTATGATGGCGACTCCTATCT -1 GAACTGGTACCAGCAGATCCCAGGGCAGCCCCCTAAGCTGCTGATCTACGACGCCTCAAATCTGGTGAGCG- GCATCCCACCACGATTCAGCGGCAGCGGCTCTGGAACC GATTTTACACTGAACATTCACCCAGTCGAGAAGGTGGACGCCGCTACCTACCATTGCCAGCAGTCTACAGA- GGACCCCTGGACTTTCGGCGGGGGAACCAAACTGGAAA TCAAG 977. 2175 L1 QSVDYDGDSY Q27-Y36 978. 2175 L1 CAGTCAGTGGACTATGATGGCGACTCCTAT -1 979. 2175 L3 QQSTEDPWT Q93- T101 980. 2175 L3 CAGCAGTCTACAGAGGACCCCTGGACT -1 981. 2175 L2 DAS D54-S56 982. 2175 L2 GACGCCTCA -1 983. 2175 VH QVQLQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFKGKATLTAD- ESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAM Q127- DYWGQGTTVTVSS S250 984. 2175 VH CAGGTGCAGCTGCAGCAGAGCGGAGCAGAGCTGGTCAGACCAGGAAGCTCCGTGAAAATTTCCTGTAAGGCAT- CTGGCTATGCCTTTTCTAGTTACTGGATGAATTGGG -1 TGAAGCAGAGGCCAGGACAGGGCCTGGAATGGATCGGGCAGATTTGGCCCGGGGATGGAGACACAAACTAT- AATGGAAAGTTCAAAGGCAAGGCTACTCTGACCGCA GACGAGTCAAGCTCCACTGCATATATGCAGCTGTCTAGTCTGGCCAGCGAGGATTCCGCTGTCTACTTTTG- CGCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTA CGCCATGGACTACTGGGGCCAGGGGACCACAGTCACCGTGTCAAGC 985. 2175 H1 GYAFSSYW G152- W159 986. 2175 H1 GGCTATGCCTTTTCTAGTTACTGG -1 987. 2175 H3 ARRETTTVGRYYYAMDY A223- Y239 988. 2175 H3 GCACGGAGAGAAACCACAACTGTGGGCAGGTACTATTACGCCATGGACTAC -1 989. 2175 H2 IWPGDGDT I177- T184 990. 2175 H2 ATTTGGCCCGGGGATGGAGACACA -1 991. 2175 CH2 APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A268- K377 992. 2175 CH2 GCTCCTGAGCTGCTGGGAGGACCAAGCGTGTTCCTGTTTCCACCTAAACCTAAGGACACTCTGATGATCTCTC-

GGACTCCCGAAGTCACCTGTGTGGTCGTGGATGTGAG -1 CCACGAGGACCCTGAAGTCAAATTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAAACAAAGC- CTAGGGAGGAACAGTATAACTCCACATACCGCGTCGTG TCTGTCCTGACTGTGCTGCATCAGGACTGGCTGAACGGAAAGGAGTACAAATGCAAGGTGAGCAACAAGGC- CCTGCCAGCTCCCATCGAGAAGACCATTTCCAAAGCTA AG 993. 2175 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G378- G483 994. 2175 CH3 GGCCAGCCTCGAGAACCACAGGTCTATGTGCTGCCACCCAGCCGGGACGAGCTGACAAAAAACCAGGTCTCCC- TGCTGTGTCTGGTGAAGGGATTCTACCCTTCTGATAT -1 TGCAGTGGAGTGGGAAAGTAATGGCCAGCCAGAAAACAATTATCTGACTTGGCCTCCAGTGCTGGATTCTG- ACGGGAGTTTCTTTCTGTACAGTAAACTGACCGTGGAT AAGTCACGGTGGCAGCAGGGAAACGTCTTTAGTTGTTCAGTGATGCACGAGGCCCTGCACAATCATTACAC- CCAGAAAAGCCTGTCCCTGTCTCCCGGC 995. 11176 Full STAILALLLAVLQGVCSQVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDG- DTNYAQKFQGRATLTADESTSTAYMELSSLRSEDTAVYYC -1 ARRETTTVGRYYYAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS- GALTSGVHTFPAVLQS5GLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKF- NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPE- NNYKTTPPVLDSDGSFALVSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG 996. 11176 Full ATGGGCTCCACCGCTATCCTGGCCCTGCTGCTGGCTGTGCTGCAGGGAGTGTGCTCCCAGGTGCAACTGGTCC- AGAGCGGCGCTGAGGTGAAGAAGCCTGGCGCCTCCG -1 TGAAGATCTCCTGCAAGGCCAGCGGCTACGCCTTCTCCTCCTACTGGATGAACTGGGTGAGGCAGGCTCCT- GGCCAGTGCCTGGAGTGGATTGGCCAGATCTGGCCCGG AGACGGCGACACCAACTACGCCCAGAAGTTCCAAGGACGGGCCACCCTGACCGCCGATGAGTCCACCTCCA- CCGCCTACATGGAGCTGTCCTCCCTGAGGTCCGAGGAC ACCGCTGTGTACTACTGTGCCAGGAGGGAGACAACCACCGTGGGCCGGTACTACTACGCCATGGACTACTG- GGGCCAGGGCACCACAGTCACCGTGTCCTCCGCCAGCA CCAAGGGCCCTTCCGTGTTCCCTCTGGCTCCCTCCTCCAAGTCCACCAGCGGAGGCACAGCCGCTCTGGGC- TGTCTGGTGAAGGACTACTTCCCCGAGCCCGTGACCGTG AGCTGGAACTCCGGAGCCCTGACCTCCGGCGTGCATACCTTCCCCGCCGTGCTGCAGTCCTCCGGACTGTA- CTCCCTGTCCTCCGTGGTGACAGTGCCCTCCTCCTCCCTG GGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTTGATAAGAAGGTGGAGCC- CAAGTCCTGCGACAAGACACACACCTGCCCTCCTTGTCC TGCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCA- GCAGGACCCCCGAGGTGACCTGCGTGGTGGTGTCCGTGA GCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAG- CCCCGGGAGGAGCAGTACAACAGCACCTACAGGGTG GTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAA- GGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCAAGG CCAAGGGCCAGCCCAGGGAACCTCAGGTGTACGTGTACCCCCCCTCCAGGGACGAGCTGACCAAGAACCAG- GTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCTC CGACATCGCTGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCTCCCGTGCTGG- ACAGCGACGGATCCTTCGCCCTGGTGTCCAAGCTGACC GTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCA- CTACACCCAGAAGTCCCTGTCCCTGTCCCCC 997. 11176 VH QVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDGDTNYAQKFQGRATLTAD- ESTSTAYMELSSLRSEDTAVYYCARRETTTVGRYYYAM Q18-S141 DYWGQGTTVTVSS 998. 11176 VH CAGGTGCAACTGGTCCAGAGCGGCGCTGAGGTGAAGAAGCCTGGCGCCTCCGTGAAGATCTCCTGCAAGGCCA- GCGGCTACGCCTTCTCCTCCTACTGGATGAACTGGG -1 TGAGGCAGGCTCCTGGCCAGTGCCTGGAGTGGATTGGCCAGATCTGGCCCGGAGACGGCGACACCAACTAC- GCCCAGAAGTTCCAAGGACGGGCCACCCTGACCGCCG ATGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGAGGTCCGAGGACACCGCTGTGTACTACTGT- GCCAGGAGGGAGACAACCACCGTGGGCCGGTACTACTA CGCCATGGACTACTGGGGCCAGGGCACCACAGTCACCGTGTCCTCC 999. 11176 H1 GYAFSSYW G43-W50 1000. 11176 H1 GGCTACGCCTTCTCCTCCTACTGG -1 1001. 11176 H3 ARRETTTVGRYYYAMDY A114- Y130 1002. 11176 H3 GCCAGGAGGGAGACAACCACCGTGGGCCGGTACTACTACGCCATGGACTAC -1 1003. 11176 H2 IWPGDGDT I68-T75 1004. 11176 H2 ATCTGGCCCGGAGACGGCGACACC -1 1005. 11176 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A142- V239 1006. 11176 CH1 GCCAGCACCAAGGGCCCTTCCGTGTTCCCTCTGGCTCCCTCCTCCAAGTCCACCAGCGGAGGCACAGCCGCTC- TGGGCTGTCTGGTGAAGGACTACTTCCCCGAGCCCGT -1 GACCGTGAGCTGGAACTCCGGAGCCCTGACCTCCGGCGTGCATACCTTCCCCGCCGTGCTGCAGTCCTCCG- GACTGTACTCCCTGTCCTCCGTGGTGACAGTGCCCTCCTC CTCCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTTGATAAGAAGG- TG 1007. 11176 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A255- K364 1008. 11176 CH2 GCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCA- GGACCCCCGAGGTGACCTGCGTGGTGGTGTCCGTGA -1 GCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAG- CCCCGGGAGGAGCAGTACAACAGCACCTACAGGGTG GTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAA- GGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCAAGG CCAAG 1009. 11176 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G365- G470 1010. 11176 CH3 GGCCAGCCCAGGGAACCTCAGGTGTACGTGTACCCCCCCTCCAGGGACGAGCTGACCAAGAACCAGGTGTCCC- TGACCTGCCTGGTGAAGGGCTTCTACCCCTCCGACA -1 TCGCTGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCTCCCGTGCTGGACAGC- GACGGATCCTTCGCCCTGGTGTCCAAGCTGACCGTGGA CAAGTCCAGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACA- CCCAGAAGTCCCTGTCCCTGTCCCCCGGC 1011. 11177 Full FGLSWLFLVAILKGVQCQVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRG- YTNYNQKVKGRFTISTDNSKNTAYLQMDSLRAEDTGVYFC -1 ARYYDDHYSLDYWGQGTLVTVSSVEGGSGGSGGSGGSGGVDDIQMTQSPSSLSASVGDRVTITCSASSSVS- YMNWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDYTL TISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT- LMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSL- LCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 1012. 11177 Full ATGGAGTTTGGCCTGTCCTGGCTGTTCCTGGTCGCTATCCTGAAGGGCGTGCAGTGCCAAGTGCAGCTGGTGC- AGTCCGGAGGAGGAGTGGTGCAGCCTGGACGGAGC -1 CTGAGGCTGTCCTGCAAGGCCTCCGGATACACCTTCACCCGGTACACCATGCATTGGGTGAGGCAGGCTCC- TGGCAAGGGCCTGGAGTGGATCGGCTACATCAATCCCA GCCGGGGATACACCAACTACAACCAGAAAGTGAAGGGCCGGTTCACCATCTCCACCGACAACAGCAAGAAC- ACCGCCTATCTGCAGATGGACTCCCTGAGGGCCGAGG ACACCGGCGTGTACTTTTGCGCTCGGTACTACGACGACCACTATTCCCTGGACTACTGGGGCCAGGGCACA- CTGGTGACAGTCTCCTCCGTGGAGGGAGGATCCGGAGG AAGCGGAGGCAGCGGAGGATCCGGCGGCGTGGATGACATCCAGATGACCCAGTCCCCTTCCTCCCTGTCCG- CTTCCGTGGGAGACAGGGTGACCATCACCTGCTCCGCC TCCAGCTCCGTGTCCTACATGAACTGGTACCAGCAAAAGCCCGGCAAGGCCCCTAAGCGGCTGATCTACGA- CACAAGCAAGCTGGCTTCCGGCGTGCCTTCCAGGTTTTC CGGCAGCGGCTCCGGCACAGACTACACCCTGACCATCAGCAGCCTGCAGCCCGAGGATGCCGCCACCTACT- ACTGCCAGCAGTGGTCCAGCAACCCTTTCACCTTCGGCC AGGGCACCAAGCTGGAGATCAAGGCCGCTGAGCCCAAGAGCTCCGACAAGACACACACCTGCCCTCCTTGT- CCTGCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCT GTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGTCCG- TGAGCCACGAGGACCCCGAGGTGAAGTTCAACTGGTAC GTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAGCCCCGGGAGGAGCAGTACAACAGCACCTACAGGGT- GGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCT GAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCA- AGGCCAAGGGCCAGCCCAGGGAACCTCAGGTGTACGT GCTGCCCCCCTCCAGGGACGAGCTGACCAAGAACCAGGTGTCCCTGCTGTGCCTGGTGAAGGGCTTCTACC- CCTCCGACATCGCTGTGGAGTGGGAGTCCAACGGCCAG CCCGAGAACAACTACCTGACCTGGCCCCCTGTGCTGGATTCCGACGGCTCCTTCTTCCTGTACTCCAAGCT- GACCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTT CTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCCC 1013. 11177 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- NSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ Q18-S136 GTLVTVSS 1014. 11177 VH CAAGTGCAGCTGGTGCAGTCCGGAGGAGGAGTGGTGCAGCCTGGACGGAGCCTGAGGCTGTCCTGCAAGGCCT- CCGGATACACCTTCACCCGGTACACCATGCATTGG -1 GTGAGGCAGGCTCCTGGCAAGGGCCTGGAGTGGATCGGCTACATCAATCCCAGCCGGGGATACACCAACTA- CAACCAGAAAGTGAAGGGCCGGTTCACCATCTCCACC GACAACAGCAAGAACACCGCCTATCTGCAGATGGACTCCCTGAGGGCCGAGGACACCGGCGTGTACTTTTG- CGCTCGGTACTACGACGACCACTATTCCCTGGACTACTG GGGCCAGGGCACACTGGTGACAGTCTCCTCC 1015. 11177 H1 GYTFTRYT G43-T50 1016. 11177 H1 GGATACACCTTCACCCGGTACACC -1 1017. 11177 H3 ARYYDDHYSLDY A114- Y125 1018. 11177 H3 GCTCGGTACTACGACGACCACTATTCCCTGGACTAC -1 1019. 11177 H2 INPSRGYT I68-T75 1020. 11177 H2 ATCAATCCCAGCCGGGGATACACC -1 1021. 11177 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDYTLT- ISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIK D155- K260 1022. 11177 VL GACATCCAGATGACCCAGTCCCCTTCCTCCCTGTCCGCTTCCGTGGGAGACAGGGTGACCATCACCTGCTCCG- CCTCCAGCTCCGTGTCCTACATGAACTGGTACCAGCAA -1 AAGCCCGGCAAGGCCCCTAAGCGGCTGATCTACGACACAAGCAAGCTGGCTTCCGGCGTGCCTTCCAGGTT- TTCCGGCAGCGGCTCCGGCACAGACTACACCCTGACCA TCAGCAGCCTGCAGCCCGAGGATGCCGCCACCTACTACTGCCAGCAGTGGTCCAGCAACCCTTTCACCTTC- GGCCAGGGCACCAAGCTGGAGATCAAG 1023. 11177 L1 SSVSY S181- Y185 1024. 11177 L1 AGCTCCGTGTCCTAC -1 1025. 11177 L3 QQWSSNPFT Q242- T250 1026. 11177 L3 CAGCAGTGGTCCAGCAACCCTTTCACC -1 1027. 11177 L2 DTS D203- S205 1028. 11177 L2 GACACAAGC -1 1029. 11177 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A278- K387 1030. 11177 CH2 GCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCA- GGACCCCCGAGGTGACCTGCGTGGTGGTGTCCGTGA -1 GCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAG- CCCCGGGAGGAGCAGTACAACAGCACCTACAGGGTG GTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAA- GGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCAAGG CCAAG 1031. 11177 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G388- G493 1032. 11177 CH3 GGCCAGCCCAGGGAACCTCAGGTGTACGTGCTGCCCCCCTCCAGGGACGAGCTGACCAAGAACCAGGTGTCCC- TGCTGTGCCTGGTGAAGGGCTTCTACCCCTCCGACA -1 TCGCTGTGGAGTGGGAGTCCAACGGCCAGCCCGAGAACAACTACCTGACCTGGCCCCCTGTGCTGGATTCC- GACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGAC AAGTCCAGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACAC- CCAGAAGTCCCTGTCCCTGTCCCCCGGC 1033. 11178 Full STAILALLLAVLQGVCSQVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDG- DTNYAQKFQGRATLTADESTSTAYMELSSLRSEDTAVYYC -1

ARRETTTVGRYYYAMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNS- GALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNH KPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKF- NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPE- NNYLTWPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPG 1034. 11178 Full ATGGGATCCACCGCTATCCTGGCTCTCCTGCTGGCTGTGCTGCAGGGCGTGTGTAGCCAGGTGCAGCTGGTGC- AGAGCGGCGCTGAAGTGAAGAAGCCCGGCGCCTCC -1 GTGAAGATCTCCTGCAAGGCCTCCGGCTACGCCTTTAGCTCCTACTGGATGAACTGGGTCAGGCAGGCTCC- CGGCCAGTGTCTCGAGTGGATCGGCCAGATTTGGCCTG GCGATGGCGACACCAACTACGCCCAGAAATTCCAGGGCAGGGCCACCCTGACCGCTGATGAGTCCACCTCC- ACCGCCTACATGGAGCTGTCCTCCCTGCGGTCCGAGGA TACCGCCGTGTACTACTGCGCCCGGAGGGAGACCACCACCGTGGGCAGGTACTACTACGCCATGGACTACT- GGGGCCAGGGCACAACCGTGACCGTGAGCTCCGCCAG CACCAAGGGACCCTCCGTGTTTCCCCTGGCTCCCTCCTCCAAGTCCACCTCCGGAGGCACAGCTGCTCTGG- GCTGTCTGGTGAAGGACTATTTTCCCGAGCCCGTGACAGT GAGCTGGAACTCCGGCGCCCTGACATCCGGCGTGCACACATTCCCTGCCGTGCTGCAGTCCTCCGGCCTGT- ACTCCCTGAGCTCCGTGGTGACAGTGCCCAGCAGCAGCC TGGGCACCCAGACCTACATCTGCAACGTGAACCACAAACCCTCCAACACAAAGGTGGACAAGAAGGTGGAG- CCCAAGTCCTGTGACAAGACACACACCTGCCCTCCTTG TCCTGCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGA- TCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGTCCG TGAGCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACC- AAGCCCCGGGAGGAGCAGTACAACAGCACCTACAGG GTGGTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAA- CAAGGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCA AGGCCAAGGGCCAGCCCAGGGAACCTCAGGTGTACGTGCTGCCCCCCTCCAGGGACGAGCTGACCAAGAAC- CAGGTGTCCCTGCTGTGCCTGGTGAAGGGCTTCTACCC CTCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACCTGACCTGGCCCCCTGTGC- TGGATTCCGACGGCTCCTTCTTCCTGTACTCCAAGCTGA CCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAAC- CACTACACCCAGAAGTCCCTGTCCCTGTCCCCC 1035. 11178 VH QVQLVQSGAEVKKPGASVKISCKASGYAFSSYWMNWVRQAPGQCLEWIGQIWPGDGDTNYAQKFQGRATLTAD- ESTSTAYMELSSLRSEDTAVYYCARRETTTVGRYYYAM Q18-S141 DYWGQGTTVTVSS 1036. 11178 VH CAGGTGCAGCTGGTGCAGAGCGGCGCTGAAGTGAAGAAGCCCGGCGCCTCCGTGAAGATCTCCTGCAAGGCCT- CCGGCTACGCCTTTAGCTCCTACTGGATGAACTGG -1 GTCAGGCAGGCTCCCGGCCAGTGTCTCGAGTGGATCGGCCAGATTTGGCCTGGCGATGGCGACACCAACTA- CGCCCAGAAATTCCAGGGCAGGGCCACCCTGACCGCT GATGAGTCCACCTCCACCGCCTACATGGAGCTGTCCTCCCTGCGGTCCGAGGATACCGCCGTGTACTACTG- CGCCCGGAGGGAGACCACCACCGTGGGCAGGTACTACT ACGCCATGGACTACTGGGGCCAGGGCACAACCGTGACCGTGAGCTCC 1037. 11178 H1 GYAFSSYW G43-W50 1038. 11178 H1 GGCTACGCCTTTAGCTCCTACTGG -1 1039. 11178 H3 ARRETTTVGRYYYAMDY A114- Y130 1040. 11178 H3 GCCCGGAGGGAGACCACCACCGTGGGCAGGTACTACTACGCCATGGACTAC -1 1041. 11178 H2 IWPGDGDT I68-T75 1042. 11178 H2 ATTTGGCCTGGCGATGGCGACACC -1 1043. 11178 CH1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS- SSLGTQTYICNVNHKPSNTKVDKKV A142- V239 1044. 11178 CH1 GCCAGCACCAAGGGACCCTCCGTGTTTCCCCTGGCTCCCTCCTCCAAGTCCACCTCCGGAGGCACAGCTGCTC- TGGGCTGTCTGGTGAAGGACTATTTTCCCGAGCCCGTG -1 ACAGTGAGCTGGAACTCCGGCGCCCTGACATCCGGCGTGCACACATTCCCTGCCGTGCTGCAGTCCTCCGG- CCTGTACTCCCTGAGCTCCGTGGTGACAGTGCCCAGCAG CAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAACCCTCCAACACAAAGGTGGACAAGAAGG- TG 1045. 11178 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A255- K364 1046. 11178 CH2 GCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCA- GGACCCCCGAGGTGACCTGCGTGGTGGTGTCCGTGA -1 GCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAG- CCCCGGGAGGAGCAGTACAACAGCACCTACAGGGTG GTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAA- GGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCAAGG CCAAG 1047. 11178 CH3 GQPREPQVYVLPPSRDELTKNQVSLLCLVKGFYPSDIAVEWESNGQPENNYLTWPPVLDSDGSFFLYSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G365- G470 1048. 11178 CH3 GGCCAGCCCAGGGAACCTCAGGTGTACGTGCTGCCCCCCTCCAGGGACGAGCTGACCAAGAACCAGGTGTCCC- TGCTGTGCCTGGTGAAGGGCTTCTACCCCTCCGACA -1 TCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACCTGACCTGGCCCCCTGTGCTGGATTCC- GACGGCTCCTTCTTCCTGTACTCCAAGCTGACCGTGGAC AAGTCCAGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACAC- CCAGAAGTCCCTGTCCCTGTCCCCCGGC 1049. 11179 Full FGLSWLFLVAILKGVQCQVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRG- YTNYNQKVKGRFTISTDNSKNTAYLQMDSLRAEDTGVYFC -1 ARYYDDHYSLDYWGQGTLVTVSSVEGGSGGSGGSGGSGGVDDIQMTQSPSSLSASVGDRVTITCSASSSVS- YMNWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDYTL TISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIKAAEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT- LMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREE QYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYVYPPSRDELTKNQVSL- TCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 1050. 11179 Full ATGGAGTTTGGCCTCTCCTGGCTGTTTCTGGTGGCTATCCTGAAGGGCGTGCAGTGCCAGGTGCAGCTCGTGC- AGTCCGGAGGAGGCGTCGTGCAACCTGGCAGGAGCC -1 TGAGGCTGTCCTGCAAGGCCAGCGGCTACACCTTCACCCGGTACACCATGCATTGGGTGAGGCAGGCCCCC- GGCAAAGGCCTGGAATGGATCGGCTACATCAACCCCTC CAGGGGCTACACCAACTACAACCAGAAGGTGAAGGGCCGGTTCACCATCTCCACCGACAACTCCAAGAACA- CAGCCTACCTGCAGATGGACTCCCTGAGGGCTGAAGAC ACAGGCGTGTATTTCTGCGCCAGGTACTACGACGACCACTACTCCCTCGACTACTGGGGACAGGGCACCCT- GGTGACCGTGTCCTCCGTGGAGGGAGGATCCGGAGGAT CCGGAGGATCCGGAGGCTCCGGAGGAGTGGACGACATCCAGATGACCCAGAGCCCTAGCTCCCTGAGCGCT- TCCGTGGGAGACCGGGTGACAATCACCTGCAGCGCCT CCTCCTCCGTGTCCTACATGAACTGGTACCAGCAGAAGCCCGGCAAGGCCCCCAAGAGGCTGATCTACGAC- ACATCCAAGCTGGCCTCCGGAGTGCCCTCCAGGTTTAGC GGCAGCGGATCCGGCACCGACTACACCCTGACCATCTCCTCCCTGCAGCCCGAGGATGCCGCTACCTATTA- CTGCCAGCAGTGGTCCAGCAACCCCTTCACCTTCGGCCA GGGCACAAAGCTGGAGATCAAGGCCGCCGAGCCCAAGTCCTCCGACAAGACACACACCTGCCCTCCTTGTC- CTGCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGT TCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCAGGACCCCCGAGGTGACCTGCGTGGTGGTGTCCGTG- AGCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGT GGACGGCGTGGAGGTGCACAACGCTAAGACCAAGCCCCGGGAGGAGCAGTACAACAGCACCTACAGGGTGG- TGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGA ACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCAAG- GCCAAGGGCCAGCCCAGGGAACCTCAGGTGTACGTGT ACCCCCCCTCCAGGGACGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCC- TCCGACATCGCCGTGGAGTGGGAGTCCAACGGCCAGCC TGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCCGACGGCTCCTTCGCCCTGGTGTCCAAGCTGA- CCGTGGACAAGTCCAGGTGGCAGCAGGGCAACGTGTTC TCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCCCCC 1051. 11179 VH QVQLVQSGGGVVQPGRSLRLSCKASGYTFTRYTMHWVRQAPGKGLEWIGYINPSRGYTNYNQKVKGRFTISTD- NSKNTAYLQMDSLRAEDTGVYFCARYYDDHYSLDYWGQ Q18-S136 GTLVTVSS 1052. 11179 VH CAGGTGCAGCTCGTGCAGTCCGGAGGAGGCGTCGTGCAACCTGGCAGGAGCCTGAGGCTGTCCTGCAAGGCCA- GCGGCTACACCTTCACCCGGTACACCATGCATTGG -1 GTGAGGCAGGCCCCCGGCAAAGGCCTGGAATGGATCGGCTACATCAACCCCTCCAGGGGCTACACCAACTA- CAACCAGAAGGTGAAGGGCCGGTTCACCATCTCCACC GACAACTCCAAGAACACAGCCTACCTGCAGATGGACTCCCTGAGGGCTGAAGACACAGGCGTGTATTTCTG- CGCCAGGTACTACGACGACCACTACTCCCTCGACTACTG GGGACAGGGCACCCTGGTGACCGTGTCCTCC 1053. 11179 H1 GYTFTRYT G43-T50 1054. 11179 H1 GGCTACACCTTCACCCGGTACACC -1 1055. 11179 H3 ARYYDDHYSLDY A114- Y125 1056. 11179 H3 GCCAGGTACTACGACGACCACTACTCCCTCGACTAC -1 1057. 11179 H2 INPSRGYT I68-T75 1058. 11179 H2 ATCAACCCCTCCAGGGGCTACACC -1 1059. 11179 VL DIQMTQSPSSLSASVGDRVTITCSASSSVSYMNWYQQKPGKAPKRLIYDTSKLASGVPSRFSGSGSGTDYTLT- ISSLQPEDAATYYCQQWSSNPFTFGQGTKLEIK D155- K260 1060. 11179 VL GACATCCAGATGACCCAGAGCCCTAGCTCCCTGAGCGCTTCCGTGGGAGACCGGGTGACAATCACCTGCAGCG- CCTCCTCCTCCGTGTCCTACATGAACTGGTACCAGCA -1 GAAGCCCGGCAAGGCCCCCAAGAGGCTGATCTACGACACATCCAAGCTGGCCTCCGGAGTGCCCTCCAGGT- TTAGCGGCAGCGGATCCGGCACCGACTACACCCTGACC ATCTCCTCCCTGCAGCCCGAGGATGCCGCTACCTATTACTGCCAGCAGTGGTCCAGCAACCCCTTCACCTT- CGGCCAGGGCACAAAGCTGGAGATCAAG 1061. 11179 L1 SSVSY S181- Y185 1062. 11179 L1 TCCTCCGTGTCCTAC -1 1063. 11179 L3 QQWSSNPFT Q242- T250 1064. 11179 L3 CAGCAGTGGTCCAGCAACCCCTTCACC -1 1065. 11179 L2 DTS D203- S205 1066. 11179 L2 GACACATCC -1 1067. 11179 CH2 APEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVSVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV- SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK A278- K387 1068. 11179 CH2 GCTCCTGAGGCTGCCGGAGGCCCTTCCGTGTTCCTGTTCCCCCCCAAGCCCAAGGACACCCTGATGATCAGCA- GGACCCCCGAGGTGACCTGCGTGGTGGTGTCCGTGA -1 GCCACGAGGACCCCGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAACGCTAAGACCAAG- CCCCGGGAGGAGCAGTACAACAGCACCTACAGGGTG GTGTCCGTGCTGACCGTGCTGCACCAGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTGTCCAACAA- GGCCCTGCCCGCTCCCATCGAGAAGACCATCTCCAAGG CCAAG 1069. 11179 CH3 GQPREPQVYVYPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFALVSKLTVD- KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG G388- G493 1070. 11179 CH3 GGCCAGCCCAGGGAACCTCAGGTGTACGTGTACCCCCCCTCCAGGGACGAGCTGACCAAGAACCAGGTGTCCC- TGACCTGCCTGGTGAAGGGCTTCTACCCCTCCGACA -1 TCGCCGTGGAGTGGGAGTCCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCTCCTGTGCTGGACTCC- GACGGCTCCTTCGCCCTGGTGTCCAAGCTGACCGTGGA CAAGTCCAGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACA- CCCAGAAGTCCCTGTCCCTGTCCCCCGGC 1071. 11175 Full DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIKRTVAAP -1 SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKA- DYEKHKVYACEVTHQGL5SPVTKSFNRGEC 1072. 11175 Full GACATTCAGCTGACCCAGAGCCCTTCCTCCCTGAGCGCCAGCGTGGGAGACCGGGCCACAATCACCTGCAGGG- CCAGCCAATCCGTGGACTACGAGGGCGACTCCTACC -1 TGAACTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACGACGCCTCCAACCTGGTGTCC- GGCATCCCTTCCAGGTTTAGCGGCTCCGGCAGCGGCAC CGATTTCACCCTGACCATCAGCAGCGTGCAGCCCGAGGACGCTGCCACCTACTACTGCCAGCAGAGCACCG- AGGACCCCTGGACCTTCGGCTGTGGCACAAAGCTGGAG ATCAAGAGGACCGTGGCCGCTCCCTCCGTGTTCATCTTCCCCCCCAGCGATGAACAGCTGAAGTCCGGCAC- AGCTTCCGTGGTGTGCCTGCTCAACAACTTCTACCCCAGG GAAGCCAAGGTGCAGTGGAAAGTTGATAACGCCCTGCAGAGCGGCAACTCCCAGGAGTCCGTGACAGAGCA- GGACAGCAAGGACTCCACCTACTCCCTGTCCTCCACCC TGACCCTGTCCAAGGCCGATTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACACACCAGGGCCTGTCC- TCCCCCGTGACCAAGTCCTTCAACAGGGGCGAG 1073. 11175 VL DIQLTQSPSSLSASVGDRATITCRASQSVDYEGDSYLNWYQQKPGKAPKLLIYDASNLVSGIPSRFSGSGSGT- DFTLTISSVQPEDAATYYCQQSTEDPWTFGCGTKLEIK D1-K111 1074. 11175 VL GACATTCAGCTGACCCAGAGCCCTTCCTCCCTGAGCGCCAGCGTGGGAGACCGGGCCACAATCACCTGCAGGG-

CCAGCCAATCCGTGGACTACGAGGGCGACTCCTACC -1 TGAACTGGTATCAGCAGAAGCCCGGCAAGGCCCCCAAGCTGCTGATCTACGACGCCTCCAACCTGGTGTCC- GGCATCCCTTCCAGGTTTAGCGGCTCCGGCAGCGGCAC CGATTTCACCCTGACCATCAGCAGCGTGCAGCCCGAGGACGCTGCCACCTACTACTGCCAGCAGAGCACCG- AGGACCCCTGGACCTTCGGCTGTGGCACAAAGCTGGAG ATCAAG 1075. 11175 L1 QSVDYEGDSY Q27-Y36 1076. 11175 L1 CAATCCGTGGACTACGAGGGCGACTCCTAC -1 1077. 11175 L3 QQSTEDPWT Q93- T101 1078. 11175 L3 CAGCAGAGCACCGAGGACCCCTGGACC -1 1079. 11175 L2 DAS D54-S56 1080. 11175 L2 GACGCCTCC -1 1081. 11175 CL RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLT- LSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC R112- C218 1082. 11175 CL AGGACCGTGGCCGCTCCCTCCGTGTTCATCTTCCCCCCCAGCGATGAACAGCTGAAGTCCGGCACAGCTTCCG- TGGTGTGCCTGCTCAACAACTTCTACCCCAGGGAAGC -1 CAAGGTGCAGTGGAAAGTTGATAACGCCCTGCAGAGCGGCAACTCCCAGGAGTCCGTGACAGAGCAGGACA- GCAAGGACTCCACCTACTCCCTGTCCTCCACCCTGACC CTGTCCAAGGCCGATTACGAGAAGCACAAGGTGTACGCCTGCGAGGTGACACACCAGGGCCTGTCCTCCCC- CGTGACCAAGTCCTTCAACAGGGGCGAGTGC

Sequence CWU 0 SQTB SEQUENCE LISTING The patent application contains a lengthy "Sequence Listing" section. A copy of the "Sequence Listing" is available in electronic form from the USPTO web site (http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20180193477A1). An electronic copy of the "Sequence Listing" will also be available from the USPTO upon request and payment of the fee set forth in 37 CFR 1.19(b)(3).

0 SQTB SEQUENCE LISTING The patent application contains a lengthy "Sequence Listing" section. A copy of the "Sequence Listing" is available in electronic form from the USPTO web site (http://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20180193477A1). An electronic copy of the "Sequence Listing" will also be available from the USPTO upon request and payment of the fee set forth in 37 CFR 1.19(b)(3).

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