Novel monoclonal antibodies that specifically bind to KAAG1 are
described. In some embodiments, the antibodies block the biological
activity of KAAG1 and are useful in composition in certain cancers, more
particularly in cancers that have increased cell surface expression of
KAAG1, such as ovarian, renal, lung, colorectal, breast, brain, and
prostate cancer, as well as melanoma. The invention also relates to cells
expressing the monoclonal antibodies and antigen binding fragments such
as humanized and chimeric antibodies. Additionally, methods of detecting
and treating cancer using the antibodies and fragments are also
disclosed.
| Inventors: |
Tremblay; Gilles Bernard; (La Prairie, CA)
; Filion; Mario; (Longueuil, CA)
; Sulea; Traian; (Kirkland, CA)
|
| Applicant: | | Name | City | State | Country | Type |
ADC THERAPEUTICS SA |
Epalinges | |
CH |
| |
|---|
| Family ID:
|
42225161
|
| Appl. No.:
|
15/833753
|
| Filed:
|
December 6, 2017 |
Related U.S. Patent Documents
| | | | |
|---|
|
| Application Number | Filing Date | Patent Number |
|---|
| | 13127439 | May 3, 2011 | 9855291 |
| | PCT/CA2009/001586 | Nov 3, 2009 | |
| | 15833753 | | |
| | 61213666 | Jun 30, 2009 | |
| | 61193184 | Nov 3, 2008 | |
|
|
| Current U.S. Class: |
1/1 |
| Current CPC Class: |
C07K 2317/76 20130101; A61K 2039/505 20130101; C07K 16/3038 20130101; C07K 2317/732 20130101; C07K 2317/24 20130101; A61P 35/02 20180101; C07K 2319/30 20130101; C07K 2317/55 20130101; A61K 31/7088 20130101; G01N 33/57449 20130101; A61P 35/00 20180101; A61P 35/04 20180101; A61P 37/04 20180101; C07K 2317/34 20130101 |
| International Class: |
A61K 31/7088 20060101 A61K031/7088; G01N 33/574 20060101 G01N033/574; C07K 16/30 20060101 C07K016/30 |
Claims
1.-21. (canceled)
22. An isolated antibody or antigen binding fragment thereof capable of
specific binding to KAAG1 (SEQ ID NO.:2) or to a KAAG1 variant having at
least 80% sequence identity with SEQ ID NO.:2 comprising a light chain
variable domain comprising a CDRL1 having an amino acid sequence at least
80% identical to the amino acid sequence set forth in SEQ ID NO.:39, a
CDRL2 having an amino acid sequence at least 80% identical to the amino
acid sequence set forth in SEQ ID NO.:40 and a CDRL3 having an amino acid
sequence at least 80% identical to the amino acid sequence set forth in
SEQ ID NO.:41 and a heavy chain variable domain comprising a CDRH1 having
an amino acid sequence at least 80% identical to the amino acid sequence
set forth in SEQ ID NO.:42, a CDRH2 having an amino acid sequence at
least 80% identical to the amino acid sequence set forth in SEQ ID NO.:43
and a CDRH3 having an amino acid sequence at least 80% identical to the
amino acid sequence set forth in SEQ ID NO.:44.
23.-108. (canceled)
109. An isolated antibody or antigen binding fragment thereof capable of
specific binding to KAAG1 (SEQ ID NO.:2) or to a variant thereof at least
80% sequence identity with SEQ ID NO.:2, the antibody or antigen binding
fragment thereof comprising: a. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:105 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:132, b. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:106 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:133, c. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:107 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:134, d. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:109 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:153, e. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:126 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:145, f. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:127 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:157, g. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:128 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:155, h. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:129 and the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:156, or; i. the 3CDRs of a light chain
variable domain defined in SEQ ID NO.:130 and the 3CDRs of a heavy chain
variable domain defined in SEQ ID NO.:151.
110.-199. (canceled)
200. The isolated antibody or antigen binding fragment thereof of claim
22, conjugated with a cytotoxic moiety.
201. The isolated antibody or antigen binding fragment thereof of claim
22, conjugated with a detectable moiety.
202. The isolated antibody or antigen binding fragment thereof of claim
109, conjugated with a cytotoxic moiety.
203. The isolated antibody or antigen binding fragment thereof of claim
109, conjugated with a detectable moiety.
204. A pharmaceutical composition comprising the isolated antibody or
antigen binding fragment thereof of claim 22 and a pharmaceutically
acceptable carrier.
205. A pharmaceutical composition comprising the isolated antibody or
antigen binding fragment thereof of claim 109 and a pharmaceutically
acceptable carrier.
206. A pharmaceutical composition comprising the isolated antibody or
antigen binding fragment thereof of claim 200 and a pharmaceutically
acceptable carrier.
207. A pharmaceutical composition comprising the isolated antibody or
antigen binding fragment thereof of claim 202 and a pharmaceutically
acceptable carrier.
208. A composition comprising the isolated antibody and antigen binding
fragment thereof of claim 201.
209. A composition comprising the isolated antibody and antigen binding
fragment thereof of claim 203.
210. A method of treating cancer comprising administering the
pharmaceutical composition of claim 204 to an individual in need.
211. A method of treating cancer comprising administering the
pharmaceutical composition of claim 205 to an individual in need.
212. A method of treating cancer comprising administering the
pharmaceutical composition of claim 206 to an individual in need.
213. A method of treating cancer comprising administering the
pharmaceutical composition of claim 207 to an individual in need.
214. A method for detecting KAAG1 or a KAAG1 variant, the method
comprising contacting a cell expressing KAAG1 or the KAAG1 variant or a
sample comprising or suspected of comprising KAAG1 or the KAAG1 variant
with the composition of claim 208.
215. A method for detecting KAAG1 or a KAAG1 variant, the method
comprising contacting a cell expressing KAAG1 or the KAAG1 variant or a
sample comprising or suspected of comprising KAAG1 or the KAAG1 variant
with the composition of claim 209.
Description
PRIORITY CLAIM
[0001] This patent application is a continuation of U.S. Ser. No.
13/127,439 filed on May 3, 2011, now U.S. Pat. No. 9,855,291 which is a
national stage filing under 35 U.S.C. .sctn. 371 of international
application PCT/CA2009/001586 filed on Nov. 3, 2009 which claimed
priority to U.S. provisional application No. 61/193,184 filed on Nov. 3,
2008 and U.S. provisional application No. 61/213,666 filed on Jun. 30,
2009. The entire contents of each of these priority applications are
incorporated herein by reference.
SEQUENCE LISTING
[0002] In accordance with 37 C.F.R. .sctn. 1.52(e) (5), a Sequence Listing
in the form of a text file (entitled "Sequence Listing," File name:
US15833753_ST25.txt, created on Jun. 12, 2018 of 6 kilobytes) is
incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
[0003] The present invention relates to monoclonal antibodies and antigen
binding fragments thereof that specifically binds to KAAG1 and their use
for treating certain diseases including diagnosing, preventing and
treating malignant tumors related to ovarian cancer. The present
invention also relates to the use of these antibodies for diagnosis,
prevention and treatment of various other cancer types.
BACKGROUND OF THE INVENTION
[0004] Among gynecologic malignancies, ovarian cancer accounts for the
highest tumor-related mortality in women in the United States (Jemal et
al., 2005). It is the fourth leading cause of cancer-related death in
women in the U.S (Menon et al., 2005). The American Cancer Society
estimated a total of 22,220 new cases in 2005 and attributed 16,210
deaths to the disease (Bonome et al., 2005). For the past 30 years, the
statistics have remained largely the same--the majority of women who
develop ovarian cancer will die of this disease (Chambers and
Vanderhyden, 2006). The disease carries a 1:70 lifetime risk and a
mortality rate of >60% (Chambers and Vanderhyden, 2006). The high
mortality rate is due to the difficulties with the early detection of
ovarian cancer when the malignancy has already spread beyond the ovary.
Indeed, >80% of patients are diagnosed with advanced staged disease
(stage III or IV) (Bonome et al., 2005). These patients have a poor
prognosis that is reflected in <45% 5-year survival rate, although 80%
to 90% will initially respond to chemotherapy (Berek et al., 2000). This
increased success compared to 20% 5-year survival rate years earlier is,
at least in part, due to the ability to optimally debulk tumor tissue
when it is confined to the ovaries, which is a significant prognostic
factor for ovarian cancer (Bristow R. E., 2000; Brown et al., 2004). In
patients who are diagnosed with early disease (stage I), the 5-yr
survival ranges from >90 (Chambers and Vanderhyden, 2006).
[0005] Ovarian cancer comprises a heterogeneous group of tumors that are
derived from the surface epithelium of the ovary or from surface
inclusions. They are classified into serous, mucinous, endometrioid,
clear cell, and Brenner (transitional) types corresponding to the
different types of epithelia in the organs of the female reproductive
tract (Shih and Kurman, 2005). Of these, serous tumors account for
.about.60% of the ovarian cancer cases diagnosed. Each histologic
subcategory is further divided into three groups: benign, intermediate
(borderline tumor or low malignancy potential (LMP)), and malignant,
reflecting their clinical behavior (Seidman et al., 2002). LMP represents
10% to 15% of tumors diagnosed as serous and is a conundrum as they
display atypical nuclear structure and metastatic behavior, yet they are
considerably less aggressive than high-grade serous tumors. The 5-year
survival for patients with LMP tumors is 95% in contrast to a <45%
survival for advanced high-grade disease over the same period (Berek et
al., 2000).
[0006] Presently, the diagnosis of ovarian cancer is accomplished, in
part, through routine analysis of the medical history of patients and by
performing physical, ultrasound and x-ray examinations, and hematological
screening. Two alternative strategies have been reported for early
hematological detection of serum biomarkers. One approach is the analysis
of serum samples by mass spectrometry to find proteins or protein
fragments of unknown identity that detect the presence or absence of
cancer (Mor et al., 2005; Kozak et al., 2003). However, this strategy is
expensive and not broadly available. Alternatively, the presence or
absence of known proteins/peptides in the serum is being detected using
antibody microarrays, ELISA, or other similar approaches. Serum testing
for a protein biomarker called CA-125 (cancer antigen-125) has long been
widely performed as a marker for ovarian cancer. However, although
ovarian cancer cells may produce an excess of these protein molecules,
there are some other cancers, including cancer of the fallopian tube or
endometrial cancer (cancer of the lining of the uterus), 60% of people
with pancreatic cancer, and 20%-25% of people with other malignancies
with elevated levels of CA-125. The CA-125 test only returns a true
positive result for about 50% of Stage I ovarian cancer patients and has
a 80% chance of returning true positive results from stage II, III, and
IV ovarian cancer patients. The other 20% of ovarian cancer patients do
not show any increase in CA-125 concentrations. In addition, an elevated
CA-125 test may indicate other benign activity not associated with
cancer, such as menstruation, pregnancy, or endometriosis. Consequently,
this test has very limited clinical application for the detection of
early stage disease when it is still treatable, exhibiting a positive
predictive value (PPV) of <10%. Even with the addition of ultrasound
screening to CA-125, the PPV only improves to around 20% (Kozak et al.,
2003). Thus, this test is not an effective screening test.
[0007] Despite improved knowledge of the etiology of the disease,
aggressive cytoreductive surgery, and modern combination chemotherapy,
there has been only little change in mortality. Poor outcomes have been
attributed to (1) lack of adequate screening tests for early disease
detection in combination with only subtle presentation of symptoms at
this stage--diagnosis is frequently being made only after progression to
later stages, at which point the peritoneal dissemination of the cancer
limits effective treatment and (2) the frequent development of resistance
to standard chemotherapeutic strategies limiting improvement in the
5-year survival rate of patients. The initial chemotherapy regimen for
ovarian cancer includes the combination of carboplatin (PARAPLATIN.TM.)
and paclitaxel (TAXOL.TM.). Years of clinical trials have proved this
combination to be most effective after effective surgery--reduces tumor
volume in about 80% of the women with newly diagnosed ovarian cancer and
40% to 50% will have complete regression--but studies continue to look
for ways to improve it. Recent abdominal infusion of chemotherapeutics to
target hard-to-reach cells in combination with intravenous delivery has
increased the effectiveness. However, severe side effects often lead to
an incomplete course of treatment. Some other chemotherapeutic agents
include doxorubicin, cisplatin, cyclophosphamide, bleomycin, etoposide,
vinblastine, topotecan hydrochloride, ifosfamide, 5-fluorouracil and
melphalan. More recently, clinical trials have demonstrated that
intraperitoneal administration of cisplatin confers a survival advantage
compared to systemic intravenous chemotherapy (Cannistra and McGuire,
2007). The excellent survival rates for women with early stage disease
receiving chemotherapy provide a strong rationale for research efforts to
develop strategies to improve the detection of ovarian cancer.
Furthermore, the discovery of new ovarian cancer-related biomarkers will
lead to the development of more effective therapeutic strategies with
minimal side effects for the future treatment of ovarian cancer.
[0008] Notwithstanding these recent advances in the understanding and the
treatment for ovarian cancer, the use of chemotherapy is invariably
associated with severe adverse reactions, which limit their use.
Consequently, the need for more specific strategies such as combining
antigen tissue specificity with the selectivity of monoclonal antibodies
should permit a significant reduction in off-target-associated side
effects. The use of monoclonal antibodies for the therapy of ovarian
cancer is beginning to emerge with an increasing number of ongoing
clinical trials (Oei et al., 2008; Nicodemus and berek, 2005). Most of
these trials have examined the use of monoclonal antibodies conjugated to
radioisotopes, such as yttrium-90, or antibodies that target tumor
antigens already identified in other cancer types. An example of this is
the use of bevacizumab, which targets vascular endothelial growth factor
(Burger, 2007). There are very few ovarian cancer specific antigens that
are currently under investigation as therapeutic targets for monoclonal
antibodies. Some examples include the use of a protein termed B7-H4
(Simon et al., 2006) and more recently folate receptor-alpha (Ebel et
al., 2007), the latter of which has recently entered Phase II clinical
trials.
[0009] Kidney associated antigen 1 (KAAG1) was originally cloned from a
cDNA library derived from a histocompatibility leukocyte antigen-B7 renal
carcinoma cell line as an antigenic peptide presented to cytotoxic T
lymphocytes (Van den Eynde et al., 1999; Genebank accession no. Q9UBP8).
The locus containing KAAG1 was found to encode two genes transcribed in
both directions on opposite strands. The sense strand was found to encode
a transcript that encodes a protein termed DCDC2. Expression studies by
these authors found that the KAAG1 antisense transcript was tumor
specific and exhibited very little expression in normal tissues whereas
the DCDC2 sense transcript was ubiquitously expressed (Van den Eynde et
al., 1999). The expression of the KAAG1 transcript in cancer, and in
particular ovarian cancer, renal cancer, lung cancer, colon cancer,
breast cancer and melanoma was disclosed in the published patent
application No. PCT/CA2007/001134. Van den Eynde et al., also observed
RNA expression in renal carcinomas, colorectal carcinomas, melanomas,
sarcomas, leukemias, brain tumors, thyroid tumors, mammary carcinomas,
prostatic carcinomas, oesophageal carcinomas, bladder tumor, lung
carcinomas and head and neck tumors. Recently, strong genetic evidence
obtained through linkage disequilibrium studies found that the
VMP/DCDC2/KAAG1 locus was associated with dyslexia (Schumacher et al.,
2006; Cope et al., 2005). One of these reports pointed to the DCDC2
marker as the culprit in dyslexic patients since the function of this
protein in cortical neuron migration was in accordance with symptoms of
these patients who often display abnormal neuronal migration and
maturation (Schumacher et al., 2006).
SUMMARY OF THE INVENTION
[0010] This invention relates to the expression of KAAG1 in tumor cells.
The invention also relates to specific anti-KAAG1 antibodies and antigen
binding fragments as well as kits useful for the treatment, detection and
diagnosis of cancer. The antibodies and antigen binding fragments may
more particularly be useful for the treatment, detection and diagnosis of
cancer where tumor cells expresses KAAG1, such as ovarian cancer, skin
cancer, renal cancer, colorectal cancer, sarcoma, leukemia, brain cancer,
cancer of the thyroid, breast cancer, prostate cancer, cancer of the
oesophagus, bladder cancer, lung cancer and head and neck cancer.
[0011] The present invention provides in one aspect thereof, an isolated
or substantially purified antibody or antigen binding fragment which may
be capable of specific binding to Kidney associated antigen 1 (KAAG1
defined in SEQ ID NO.:2) or to a KAAG1 variant.
[0012] More specifically and in accordance with an embodiment of the
invention, the antibody or antigen binding fragment may bind to a domain
located between amino acid 30 and amino acid 84 of KAAG1.
[0013] In accordance with another embodiment of the invention, the
antibody or antigen binding fragment may be capable of binding to an
epitope comprised within amino acid 1 to 35 of KAAG1.
[0014] In accordance with a further embodiment of the invention, the
antibody or antigen binding fragment may be capable of binding to an
epitope comprised within amino acid 36 to 60 of KAAG1.
[0015] In accordance with yet a further embodiment of the invention, the
antibody or antigen binding fragment may be capable of binding to an
epitope comprised within amino acid 61 to 84 of KAAG1.
[0016] The antibody or antigen binding fragment of the present invention
is especially capable of specific binding to a secreted form of KAAG1,
i.e., a form of KAAG1 where the signal peptide has been cleaved.
[0017] The antibody or antigen binding fragment of the present invention
is especially capable of binding to the extracellular region of KAAG1.
[0018] As such, the present invention encompasses diagnostic and/or
therapeutic antibodies or antigen binding fragments having specificity
for a secreted form of KAAG1 or for an extracellular region of KAAG1.
Also encompassed by the present invention are antibodies or antigen
binding fragments having the same epitope specificity as the antibody of
the present invention. A candidate antibody may be identified by
determining whether it will bind to the epitope to which the antibodies
described herein binds and/or by performing competition assays with
antibodies or antigen binding fragments known to bind to the epitope.
[0019] Therefore another aspect the present invention provides an isolated
antibody or antigen binding fragment capable of competing with the
antibody or antigen binding fragment described herein.
[0020] Isolated antibodies or antigen binding fragments of the present
invention include those which may be capable of inducing killing
(elimination, destruction, lysis) of KAAG1-expressing tumor cells or
KAAG1 variant-expressing tumor cells (e.g., in an ADCC-dependent manner).
[0021] Isolated antibodies or antigen binding fragments of the present
invention also include those which are characterized by their ability to
reduce spreading of KAAG1-expressing tumor cells and also those which are
characterized by their ability to decrease or impair formation of
KAAG1-expressing tumors.
[0022] The antibodies or antigen binding fragments may be particularly
effective when KAAG1 is expressed at the surface of the KAAG1-expressing
tumor cells and may be particularly useful in targeting KAAG1-expressing
tumor cells characterized by anchorage-independent growth.
[0023] The invention relates to monoclonal antibodies, polyclonal
antibodies, chimeric antibodies, humanized antibodies and human
antibodies (isolated) as well as antigen binding fragments having the
characteristics described herein. Antibodies or antigen binding fragments
encompassing permutations of the light and/or heavy chains between a
monoclonal, chimeric, humanized or human antibody are also encompassed
herewith.
[0024] The antibodies or antigen binding fragments of the present
invention may thus comprise amino acids of a human constant region and/or
framework amino acids of a human antibody.
[0025] The term "antibody" refers to intact antibody, monoclonal or
polyclonal antibodies. The term "antibody" also encompasses multispecific
antibodies such as bispecific antibodies. Human antibodies are usually
made of two light chains and two heavy chains each comprising variable
regions and constant regions. The light chain variable region comprises 3
CDRs, identified herein as CDRL1, CDRL2 and CDRL3 flanked by framework
regions. The heavy chain variable region comprises 3 CDRs, identified
herein as CDRH1, CDRH2 and CDRH3 flanked by framework regions.
[0026] The term "antigen-binding fragment", as used herein, refers to one
or more fragments of an antibody that retain the ability to bind to an
antigen (e.g., KAAG1, secreted form of KAAG1 or variants thereof). It has
been shown that the antigen-binding function of an antibody can be
performed by fragments of an intact antibody. Examples of binding
fragments encompassed within the term "antigen-binding fragment" of an
antibody include (i) a Fab fragment, a monovalent fragment consisting of
the V.sub.L, V.sub.H, C.sub.L and C.sub.H1 domains; (ii) a F(ab').sub.2
fragment, a bivalent fragment comprising two Fab fragments linked by a
disulfide bridge at the hinge region; (iii) a Fd fragment consisting of
the V.sub.H and C.sub.H1 domains; (iv) a Fv fragment consisting of the
V.sub.L and V.sub.H domains of a single arm of an antibody, (v) a dAb
fragment (Ward et al., (1989) Nature 341:544-546), which consists of a
V.sub.H domain; and (vi) an isolated complementarity determining region
(CDR), e.g., V.sub.H CDR3. Furthermore, although the two domains of the
Fv fragment, V.sub.L and V.sub.H, are coded for by separate genes, they
can be joined, using recombinant methods, by a synthetic linker that
enables them to be made as a single polypeptide chain in which the
V.sub.L and V.sub.H regions pair to form monovalent molecules (known as
single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426;
and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883). Such
single chain antibodies are also intended to be encompassed within the
term "antigen-binding fragment" of an antibody. Furthermore, the
antigen-binding fragments include binding-domain immunoglobulin fusion
proteins comprising (i) a binding domain polypeptide (such as a heavy
chain variable region, a light chain variable region, or a heavy chain
variable region fused to a light chain variable region via a linker
peptide) that is fused to an immunoglobulin hinge region polypeptide,
(ii) an immunoglobulin heavy chain CH2 constant region fused to the hinge
region, and (iii) an immunoglobulin heavy chain CH3 constant region fused
to the CH2 constant region. The hinge region may be modified by replacing
one or more cysteine residues with serine residues so as to prevent
dimerization. Such binding-domain immunoglobulin fusion proteins are
further disclosed in US 2003/0118592 and US 2003/0133939. These antibody
fragments are obtained using conventional techniques known to those with
skill in the art, and the fragments are screened for utility in the same
manner as are intact antibodies.
[0027] A typical antigen binding site is comprised of the variable regions
formed by the pairing of a light chain immunoglobulin and a heavy chain
immunoglobulin. The structure of the antibody variable regions is very
consistent and exhibits very similar structures. These variable regions
are typically comprised of relatively homologous framework regions (FR)
interspaced with three hypervariable regions termed Complementarity
Determining Regions (CDRs). The overall binding activity of the antigen
binding fragment is often dictated by the sequence of the CDRs. The FRs
often play a role in the proper positioning and alignment in three
dimensions of the CDRs for optimal antigen binding.
[0028] Antibodies and/or antigen binding fragments of the present
invention may originate, for example, from a mouse, a rat or any other
mammal or from other sources such as through recombinant DNA
technologies.
[0029] Further scope, applicability and advantages of the present
invention will become apparent from the non-restrictive detailed
description given hereinafter. It should be understood, however, that
this detailed description, while indicating exemplary embodiments of the
invention, is given by way of example only, with reference to the
accompanying drawings.
BRIEF DESCRIPTION OF THE DRAWINGS
[0030] FIG. 1A shows the expression profiling analyses using
semi-quantitative RT-PCR reactions carried out to measure the level of
KAAG1 mRNA expression in RNA samples derived from greater than 20 ovarian
tumors, benign (low malignancy potential) tumors, ovarian cancer cell
lines, and 30 normal tissues. The control panels show GAPDH expression, a
house-keeping gene used to compare the amount of starting material in
each RT-PCR reaction.
[0031] FIG. 1B shows semi-quantitative RT-PCR experiments demonstrating
that KAAG1 mRNA is expressed in ovarian cancer cell lines, in particular
those that are derived from ascites.
[0032] FIG. 1C shows a diagram illustrating the ability of ovarian cancer
cell lines to form 3D structures called spheroids. The left panels show
the cells grown in medium lacking serum whereas 5% serum stimulated the
formation of the spheroid structures.
[0033] FIG. 1D shows semi-quantitative RT-PCR experiments demonstrating
that the KAAG1 mRNA is highly induced during the formation of spheroids
in ovarian cancer cell lines.
[0034] FIG. 2A shows a diagram illustrating the wound or scratch assay, a
cell-based assay that is a measurement of a cell line's ability to
migrate into a denuded area over a pre-determined period of time. TOV-21G
cells harboring KAAG1 shRNAs display a reduced capacity to fill in the
denuded area.
[0035] FIG. 2B shows an illustration of the clonogenic assay, also known
as a colony survival assay. It measured the survival of diluted cells
over a period of several days. TOV-21G cells harboring KAAG1 shRNAs
display reduced survival.
[0036] FIG. 3A shows a polyacrylamide gel that was stained with Coomassie
Blue and contains a sample (10 .mu.g) of purified Fc-KAAG1 fusion protein
that was produced in transiently transfected 293E cells.
[0037] FIG. 3B shows the results of an ELISA of one of the 96-well plates
containing individual monoclonal antibodies selected from OMNICLONAL.TM.
library #3 containing anti-KAAG1 Fabs. The results showed that 48
(highlighted in grey) of the Fabs interacted very efficiently with KAAG1.
The wells indicated by bold numbers contained the exemplary monoclonals
3D3, 3G10, and 3C4.
[0038] FIG. 4A shows a polyacrylamide gel that was stained with Coomassie
Blue and contains a sample (10 .mu.g) of purified Fc-KAAG1 fusion protein
(lane 1), a truncated mutant of KAAG1 spanning amino acids 1-60 (lane 2),
and another truncated mutant of KAAG1 spanning amino acids 1-35 (lane 3)
that were produced in transiently transfected 293E cells. All proteins
were Fc fusion proteins.
[0039] FIG. 4B is a scheme that illustrates the truncated mutants of KAAG1
that were generated for the epitope mapping studies.
[0040] FIG. 4C shows a drawing that describes the results from ELISA
analyses to map the epitopes that are bound by the anti-KAAG1 antibodies
contained in OMNICLONAL.TM. library #3. The results showed that the
majority of monoclonals interact with central region of KAAG1 and that
certain antibodies bound to the amino- or carboxyl-termini of KAAG1.
[0041] FIG. 5 presents a scheme that illustrates the steps involved to
convert the mouse Fabs into IgG1 mouse-human chimeric mAbs.
[0042] FIG. 6 shows drawings that compare the binding of the mouse
anti-KAAG1 Fabs with the binding of the corresponding IgG1 chimeric
monoclonal antibodies for exemplary antibodies 3D3, 3G10, and 3C4. The
results indicate that the relative binding of the Fab variable regions
was maintained when transferred to a full human IgG1 scaffold.
[0043] FIG. 7 shows depictions of spheroid formation experiments using
TOV-21G and OV-90 ovarian cancer cell lines in the presence of chimeric
IgG1 anti-KAAG1 monoclonal antibodies. Loosely packed structures are
indicative of less invasive cancer cell lines. The results show spheroids
treated with the exemplary anti-KAAG1 antibodies 3D3, 3G10, or 3C4.
[0044] FIG. 8A shows a scan of a tissue microarray containing
approximately 70 biopsy samples obtained from ovarian tumor patients. The
samples were blotted with the 3D3 anti-KAAG1 antibody and showed that the
vast majority of ovarian tumors expressed very high level of KAAG1
antigen.
[0045] FIG. 8B a higher magnification picture from the tissue microarray
experiment. The arrows show the membrane localization of KAAG1 at the
apical surface of the epithelial layer of cells in serous ovarian tumors.
[0046] FIG. 8C illustrates other immunohistochemical studies that
demonstrate that KAAG1 is highly expressed in all ovarian cancer types.
The histotypes shown are serous, mucinous and endometroid.
[0047] FIGS. 9A, 9B and 9C is a summary of alignment results obtained for
selected CDRL1, CDRL2 or CDRL3 sequences using the ClustalW2 program;
where "*" means that the residues in that column are identical in all
sequences in the alignment, ":" means that conserved substitutions have
been observed and "." means that semi-conserved substitutions are
observed. Consensus CDRs were generated using the ClustalW program
(Larkin M. A., et al., (2007) ClustalW and ClustalX version 2.
Bioinformatics 2007 23(21): 2947-2948).
[0048] FIGS. 10A, 10B and 10C is a summary of alignment results obtained
for selected CDRH1, CDRH2 or CDRH3 sequences using the ClustalW2 program;
where "*" means that the residues in that column are identical in all
sequences in the alignment, ":" means that conserved substitutions have
been observed and "." means that semi-conserved substitutions are
observed. Consensus CDRs were generated using the ClustalW program
(Larkin M. A., et al., (2007) ClustalW and ClustalX version 2.
Bioinformatics 2007 23(21): 2947-2948).
[0049] FIG. 11 represents sequence comparison between each of the light
chain variable regions generated and representative light chain variable
regions identified in SEQ ID NOs:16, 20, 24 or 105. Percent sequence
identity and percent sequence similarity has been determined using Blast2
sequence program as indicated herein.
[0050] FIG. 12 represents sequence comparison between each of the heavy
chain variable regions generated and representative heavy chain variable
regions identified in SEQ ID NOs:18, 22, 26 or 132. Percent sequence
identity and percent sequence similarity has been determined using Blast2
sequence program as indicated herein.
[0051] FIG. 13 An IgG.sub.1 antibody that targets KAAG1 can efficiently
mediate ADCC activity in vitro. PBMNCs (AllCells, LLC, Emoryville,
Calif.) were incubated with 3D3 for 30 min and mixed with either OVCAR-3
or WIL2-S cells at a ratio of 1:25. The cells were incubated for 4 h at
37 C and cell lysis was determined by measuring LDH levels in the medium.
Cell cytotoxicity was calculated as follows: %
cytotoxicity=(experimental-effector spontaneous-target
spontaneous).times.100/(target maximum-target spontaneous).
[0052] FIG. 14A is a diagram representing the number of micro-metastatic
tumors in mice treated with the 3C4 or the 3D3 anti-KAAG1 antibodies in
comparison with untreated mice (PBS). The data are expressed as the
average number of tumors/mouse.+-.SE. FIG. 14B are pictures illustrating
the number of tumors visually scored in the experiment of FIG. 14A.
[0053] FIG. 15 shows immunohistochemistry performed with an anti-KAAG1
antibody on human skin tumor tissue microarrays (Pantomics Inc.,
Richmond, Calif.) of several sections isolated from squamous cell
carcinomas and melanomas.
[0054] FIG. 16 illustrates spheroid formation of melanoma cell lines (A375
and SK-MEL5) and of renal cell carcinoma cell lines (A498 and 786-O) in
the presence or absence of the chimeric 3D3 antibody.
[0055] FIG. 17A represents graphs illustrating the binding of increasing
concentrations of the 3C4, 3D3 and 3G10 antibodies to cell lines (OV-90,
TOV-21G and SKOV-3) fixed under condition that do not permeate the cells.
[0056] FIG. 17B is a graph illustrating the results of flow cytometry
performed on SKOV-3 cell line with the 3D3 antibody.
[0057] FIG. 18A is a schematic illustrating the structure of the 3D3
antibody model.
[0058] FIG. 18B is a schematic illustrating the structure of the 3C4
antibody model.
[0059] FIG. 19A is a graph illustrating the binding of increasing
concentration of the humanized 3D3 antibody in comparison with the
chimeric 3D3 antibody to recombinant KAAG1.
[0060] FIG. 19B is a table summarizing the kinetics parameters of the
humanized 3D3 antibody, the chimeric 3D3 antibody as well as hybrid
antibodies encompassing permutations of the light and heavy chains of the
chimeric or humanized antibody.
[0061] FIG. 19C illustrates spheroid formation of SKOV-3 ovarian cancer
cells in the presence of the humanized 3D3 antibody, chimeric 3D3
antibody or in the presence of a buffer or a control IgG.
[0062] FIG. 20A represents sequence alignment of the monoclonal 3D3 light
chain variable region (SEQ ID NO.:16) and the humanized 3D3 light chain
variable region (SEQ ID NO.:178). The humanized 3D3 light chain variable
region is 86% identical (94% sequence similarity) to the monoclonal 3D3
light chain variable region and their three CDRs are 100% (indicated in
bold).
[0063] FIG. 20B represents sequence alignment of the monoclonal 3D3 heavy
chain variable region (SEQ ID NO.:18) and the humanized 3D3 heavy chain
variable region (SEQ ID NO.:179). The humanized 3D3 heavy chain variable
region is 82% identical (91% sequence similarity) to the monoclonal 3D3
heavy chain variable region and their three CDRs are 100% (indicated in
bold).
[0064] FIG. 21A represents sequence alignment of the monoclonal 3C4 light
chain variable region (SEQ ID NO.:24) and the humanized 3C4 light chain
variable region (SEQ ID NO.:182). The humanized 3C4 light chain variable
region is 85% identical (93% sequence similarity) to the monoclonal 3C4
light chain variable region and their three CDRs are 100% (indicated in
bold).
[0065] FIG. 21B represents sequence alignment of the monoclonal 3C4 heavy
chain variable region (SEQ ID NO.:26) and the humanized 3C4 heavy chain
variable region (SEQ ID NO.:183). The humanized 3C4 heavy chain variable
region is 86% identical (93% sequence similarity) to the monoclonal 3C4
heavy chain variable region and their three CDRs are 100% (indicated in
bold).
DETAILED DESCRIPTION OF THE INVENTION
The Expression and Biological Activity of KAAG1 in Cancer Cells
[0066] The present invention relates to the use of antibodies to target
tumors found in various cancer types, in particular ovarian cancer. In
order to direct the antibodies to the tumors, the identification of
tumor-specific antigens that are expressed at the cell surface of the
cancer cells must be carried out. There are several technologies that are
available to identify tumor-specific antigens and the method that was
used to identify KAAG1 in ovarian tumors, an innovative discovery
platform called Subtractive Transcription-based Amplification of mRNA
(STAR), is described in the published patent application No.
PCT/CA2007/001134.
[0067] Analysis of the ovarian cancer STAR libraries yielded many genes
that encode secreted and cell surface proteins. One of these, termed
AB-0447, contained an open reading frame that encoded a polypeptide of 84
amino acids, corresponding to SEQ ID NO.:2 that was encoded by a cDNA of
885 base pairs with the nucleotide sequence shown in SEQ ID NO.:1. A
search of publicly available databases revealed that the AB-0447
nucleotide sequence was identical to that of a gene called KAAG1.
Bioinformatic analysis predicted a membrane-anchored protein that
presents its functional domain to the extracellular compartment. KAAG1
was originally cloned from a kidney cancer library as a cell surface
antigen, a result that confirms its membrane localization. Additionally,
our studies showed that the protein was processed at its amino-terminus,
a result that was consistent with cleavage of a functional signal peptide
at or between amino acids 30 and 34. Furthermore, transient expression of
the full-length cDNA resulted in detection of cleaved KAAG1 in the
culture medium. This last finding indicated that this membrane-anchored
protein could be shed from the cells when expressed at high levels. In
contrast, expression of an amino-truncated mutant of KAAG1 resulted in
intra-cellular retention of the protein. There are currently no published
reports that shed any light on its function and the over-expression of
KAAG1 in ovarian cancer, as disclosed by this invention, has never been
previously documented.
[0068] We have thus investigated whether KAAG1 could be used for
antibody-based diagnostics and therapeutics.
[0069] Several ovarian cancer cell-based models have been established,
such as TOV-21G, TOV-112D, OV-90, and others, and are familiar to those
skilled in the art. These cells are part of a collection of human ovarian
cancer cell lines derived from patients with ovarian tumors or ascites
fluid. These cell lines have undergone an in-depth analysis, including
global gene expression patterns on microarrays that make them excellent
cell-based models for human ovarian cancer. The growth properties, gene
expression patterns, and response to chemotherapeutic drugs indicated
that these cell lines are very representative of ovarian tumor behavior
in vivo (Benoit et al., 2007). RT-PCR analysis of total RNA isolated from
these ovarian cancer cell lines showed that the KAAG1 transcript was
weakly expressed in the cell lines derived from primary tumors. In
contrast, cell lines derived from ascitic fluid contained high levels of
KAAG1 expression. The increased expression of KAAG1 in cells from the
ascitic fluid suggested that the environment of the cells influences the
regulation of the KAAG1 gene. Ascitic cells are associated with advanced
disease and this pattern of expression implies that increased KAAG1
levels are associated with anchorage-independent growth. In concordance
with this latter suggestion, KAAG1 expression was found to significantly
increase in cell lines derived from primary tumors when these cells were
cultured as spheroids in 3D cultures. These spheroids have been
extensively characterized and were found to display many properties
associated with tumors in vivo (Cody et al., 2008). Thus, expression of
KAAG1 was found to be significantly increased in models that mimic tumor
progression, in particular during the evolution of ovarian cancer.
[0070] With the demonstration that KAAG1 expression is regulated in
ovarian cancer cells, the function of this gene in ovarian cancer cell
behavior was examined in cell-based assays. To that effect, RNA
interference (RNAi) was used to knock down the expression of the
endogenous KAAG1 gene in the ovarian cancer cell lines and it was found
that decreased expression of KAAG1 resulted in a significant reduction in
the migration of the cells as determined in a standard cell motility
assay, as exemplified by a wound healing (or scratch) assay. This type of
assay measures the speed at which cells fill a denuded area in a
confluent monolayer. Decreased expression of KAAG1 resulted in a
reduction in the survival of ovarian cancer cell lines as measured by a
clonogenic assay, such as a colony survival assay. Those skilled in the
art may use other methods to evaluate the requirement of KAAG1 in the
behavior of cancer cells, in particular ovarian cancer cells.
[0071] Based on the expression of KAAG1 in a large proportion of ovarian
tumors, its limited expression in normal tissues, and a concordance
between expression levels and increased malignancy, and a putative
biological role for KAAG1 in the behavior of ovarian cancer cell lines,
KAAG1 was chosen as a therapeutic target for the development of
antibodies for the detection, prevention, and treatment of ovarian
cancer. Expression of KAAG1 in cancer, other than ovarian cancer also
lead the Applicant to the evaluation of therapeutic or diagnostic
antibodies for other cancer indications.
[0072] Therefore, a variety of anti-KAAG1 antibodies and antigen binding
fragments thereof, such as monoclonal antibodies, polyclonal antibodies,
chimeric and humanized antibodies (including humanized monoclonal
antibodies), antibody fragments, single chain antibodies, domain
antibodies, and polypeptides with an antigen binding region, useful for
targeting KAAG1 are provided.
KAAG1 as Antigen and Epitopes Derived from KAAG1
[0073] The Applicant has come to the unexpected discovery that KAAG1 is
expressed in several tumor types and is also found in blood and in
ascitic fluid of patients. This antigen may thus be useful for targeting
tumor cells expressing the antigen in vivo and in the development of
detection assays for measuring the tumor associated antigen in vitro or
in vivo. The KAAG1 antigen circulating in blood lacks the signal peptide.
[0074] The present invention therefore provides a KAAG1 antigen useful for
generating antibodies specific for the circulating form of KAAG1 and/or
specific for tumor-expressed KAAG1. The KAAG1 antigen (i.e., epitope) may
comprise a fragment of at least 10 amino acids (and up to 84 amino acids)
of KAAG1 and may especially bind to the extracellular region of KAAG1.
[0075] An exemplary antigen is the whole KAAG1 protein or a variant form
having at least 80% sequence identity with SEQ ID NO.:2 or a fragment
thereof.
[0076] Another exemplary antigen derived from KAAG1 is the secreted or
circulating form of KAAG1 which lacks the signal peptide or the
extracellular region of KAAG1. This antigen may more particularly lack
amino acids 1 to 25, 1 to 26, 1 to 27, 1 to 28, 1 to 29, 1 to 30, 1 to
31, 1 to 32, 1 to 33, 1 to 34, 1 to 35 or 1 to 36 of KAAG1.
[0077] The antigen or the epitope described herein may be fused with a
carrier such as keyhole limpet (KHL), bovine serum albumin (BSA),
ovalbumin (OVA) or else in order to generate antibodies and antigen
binding fragments.
[0078] The present invention also provides an epitope comprised within
amino acid 1 to 35 of SEQ ID NO.:2, within amino acid 36 to 60 of SEQ ID
NO.:2 or within amino acid 61 to 84 of SEQ ID NO.:2 to generate
antibodies and antigen binding fragments described herein. The present
invention further provides a composition for generating antibodies to a
secreted or circulating form of KAAG1 or to an extracellular region of
KAAG1, the composition may comprise an epitope of KAAG1 comprised within
amino acids 30 to 84 of SEQ ID NO.:2 and a carrier. The epitope may
especially comprise at least 10 amino acids of KAAG1.
[0079] Exemplary embodiments of compositions are pharmaceutical
composition for generating antibodies to a secreted or circulating form
of KAAG1 or to the extracellular region of KAAG1. The pharmaceutical
composition may comprise an epitope of KAAG1 comprised within amino acids
30 to 84 of SEQ ID NO.:2 and a pharmaceutically acceptable carrier.
[0080] In yet a further aspect the invention provides a method for
generating antibodies to a secreted or circulating form of KAAG1. The
method may comprise administering a polypeptide comprising an epitope of
KAAG1 comprised within amino acids 30 to 84 of SEQ ID NO.:2 wherein the
epitope lacks a KAAG1 signal peptide.
[0081] Alternatively, the method may comprise administering an epitope
which comprises the signal peptide and selecting antibodies which only
binds to the secreted form or the extracellular region of the protein.
[0082] In an additional aspect, the present invention provides the use of
an epitope of KAAG1 comprised within amino acids 30 to 84 of SEQ ID NO.:2
for generating antibodies to a secreted or circulating form of KAAG1.
Antibodies and Antigen Binding Fragments that Binds to KAAG1
[0083] Antibodies were initially isolated from Fab libraries for their
specificity towards the antigen of interest. Comparison of the amino acid
sequences of the light chain variable domains or the heavy chain variable
domains of antibodies showing the greatest characteristics allowed us to
derive consensus sequences within the CDRs and within the variable
regions. The consensus for CDRs are provided in SEQ ID Nos: 74 to 90.
[0084] The variable regions described herein may be fused with constant
regions of a desired species thereby allowing recognition of the antibody
by effector cells of the desired species. The constant region may
originate, for example, from an IgG1, IgG2, IgG3, or IgG4 subtype.
Cloning or synthesizing a constant region in frame with a variable region
is well within the scope of a person of skill in the art and may be
performed, for example, by recombinant DNA technology.
[0085] In certain embodiments of the present invention, antibodies that
bind to KAAG1 may be of the IgG1, IgG2, IgG3, or IgG4 subtype. More
specific embodiments of the invention relates to an antibody of the IgG1
subtype. The antibody may be a humanized antibody of the IgG1 subtype
that is biologically active in mediating antibody-dependent cellular
cytotoxicity (ADCC), complement-mediated cytotoxicity (CMC), or
associated with immune complexes. The typical ADCC involves activation of
natural killer (NK) cells and is reliant on the recognition of
antibody-coated cells by Fc receptors on the surface of the NK cells. The
Fc receptors recognize the Fc domain of antibodies such as is present on
IgG1, which bind to the surface of a target cell, in particular a
cancerous cell that expresses an antigen, such as KAAG1. Once bound to
the Fc receptor of IgG the NK cell releases cytokines and cytotoxic
granules that enter the target cell and promote cell death by triggering
apoptosis.
[0086] In some instances, anti-KAAG1 antibodies with substantially
identical light and heavy chain variable regions to antibody 3D3, will
interact with an epitope spanned by amino acids 36-60, inclusively, of
KAAG1. In other instances, anti-KAAG1 antibodies with substantially
identical light and heavy chain variable regions to antibody 3G10, will
interact with an epitope spanned by amino acids 61-84, inclusively, of
KAAG1. In yet another instance, anti-KAAG1 antibodies with substantially
identical light and heavy chain variable regions to antibody 3C4 will
interact with an epitope spanned by amino acids 1-35, inclusively, of
KAAG1.
[0087] The present invention described a collection of antibodies that
bind to KAAG1. In certain embodiments, the antibodies may be selected
from the group consisting of polyclonal antibodies, monoclonal antibodies
such as chimeric or humanized antibodies, antibody fragments such as
antigen binding fragments, single chain antibodies, domain antibodies,
and polypeptides with an antigen binding region.
[0088] In an aspect of the invention, the isolated antibody or antigen
binding fragment of the present invention may be capable of inducing
killing (elimination, destruction, lysis) of KAAG1-expressing tumor cells
or KAAG1 variant-expressing tumor cells (e.g., in an ADCC-dependent
manner).
[0089] In a further aspect of the invention, the isolated antibody or
antigen binding fragment of the present invention may especially be
characterized by its capacity of reducing spreading of KAAG1-expressing
tumor cells.
[0090] In an additional aspect of the invention, the isolated antibody or
antigen binding fragment of the present invention may be characterized by
its capacity of decreasing or impairing formation of KAAG1-expressing
tumors.
[0091] In accordance with an embodiment of the invention, the antibody or
antigen binding fragment may be more particularly effective when KAAG1 is
expressed at the surface of the KAAG1-expressing tumor cells.
[0092] Also in accordance with the present invention, the antibody or
antigen binding fragment may be especially useful in targeting
KAAG1-expressing tumor cells which are characterized by
anchorage-independent growth.
[0093] In a further aspect, the present invention relates to an isolated
antibody or antigen binding fragment for use in the treatment of cancer
comprising tumor cells expressing KAAG1.
[0094] In yet a further aspect, the present invention relates to an
isolated antibody or antigen binding fragment for use in the detection of
cancer comprising tumor cells expressing KAAG1.
[0095] In an exemplary embodiment of the invention, the isolated antibody
or antigen binding fragment may comprise amino acids of a constant
region, which may originate, for example, from a human antibody.
[0096] In another exemplary embodiment of the invention, the isolated
antibody or antigen binding fragment may comprise framework amino acids
of a human antibody.
[0097] Without being limited to the exemplary embodiments presented
herein, the Applicant as generated specific antibodies and antigen
binding fragments which may be useful for the purposes described herein.
[0098] The present invention therefore provides in an exemplary
embodiment, an isolated antibody or antigen binding fragment comprising a
light chain variable domain having; [0099] a. a CDRL1 sequence selected
from the group consisting of SEQ ID NO.:74 and SEQ ID NO.:75; [0100] b. a
CDRL2 sequence selected from the group consisting of SEQ ID NO.:76, SEQ
ID NO.: 77 and SEQ ID NO.:78, or; [0101] c. a CDRL3 sequence selected
from the group consisting of SEQ ID NO.:79, SEQ ID NO.:80 and SEQ ID
NO.:81.
[0102] The isolated antibody or antigen binding fragment may also comprise
a heavy chain variable domain having; [0103] a. a CDRH1 sequence
comprising SEQ ID NO.:82; [0104] b. a CDRH2 sequence selected from the
group consisting of SEQ ID NO.:83, SEQ ID NO.:84, SEQ ID NO.:85, SEQ ID
NO.:86 and SEQ ID NO.:87, or; [0105] c. a CDRH3 sequence selected from
the group consisting of SEQ ID NO.:88, SEQ ID NO.:89 and SEQ ID NO.:90.
[0106] In an exemplary embodiment, the antibody or antigen binding
fragment may comprise any individual CDR or a combination of CDR1, CDR2
and/or CDR3 of the light chain variable region. The CDR3 may more
particularly be selected. Combination may include for example, CDRL1 and
CDRL3; CDRL1 and CDRL2; CDRL2 and CDRL3 and; CDRL1, CDRL2 and CDRL3.
[0107] In another exemplary embodiment, the antibody or antigen binding
fragment may comprise any individual CDR or a combination of CDR1, CDR2
and/or CDR3 of the heavy chain variable region. The CDR3 may more
particularly be selected. Combination may include for example, CDRH1 and
CDRH3; CDRH1 and CDRH2; CDRH2 and CDRH3 and; CDRH1, CDRH2 and CDRH3.
[0108] In accordance with the present invention, the antibody or antigen
binding fragment may comprise at least two CDRs of a CDRL1, a CDRL2 or a
CDRL3.
[0109] Also in accordance with the present invention, the antibody or
antigen binding fragment may comprise one CDRL1, one CDRL2 and one CDRL3.
[0110] Further in accordance with the present invention, the antibody or
antigen binding fragment may comprise: [0111] a. At least two CDRs of a
CDRL1, CDRL2 or CDRL3 and; [0112] b. At least two CDRs of a CDRH1, one
CDRH2 or one CDRH3.
[0113] The antibody or antigen binding fragment may more preferably
comprise one CDRL1, one CDRL2 and one CDRL3.
[0114] The antibody or antigen binding fragment may also more preferably
comprise one CDRH1, one CDRH2 and one CDRH3.
[0115] Other exemplary embodiments of the invention relates to an isolated
antibody or antigen binding fragment comprising a heavy chain variable
domain having; [0116] a. a CDRH1 sequence comprising SEQ ID NO.:82;
[0117] b. a CDRH2 sequence selected from the group consisting of SEQ ID
NO.:83, SEQ ID NO.:84, SEQ ID NO.:85, SEQ ID NO.:86 and SEQ ID NO.:87,
or; [0118] c. a CDRH3 sequence selected from the group consisting of SEQ
ID NO.:88, SEQ ID NO.:89 and SEQ ID NO.:90.
[0119] In accordance with the present invention, the antibody or antigen
binding fragment may comprise one CDRH1, one CDRH2 or one CDRH3.
[0120] In accordance with the present invention, the antibody or antigen
binding fragment may also comprise one CDRH1, one CDRH2 and one CDRH3.
[0121] When only one of the light chain variable domain or the heavy chain
variable domain is available, an antibody or antigen-binding fragment may
be reconstituted by screening a library of complementary variable domains
using methods known in the art (Portolano et al. The Journal of
Immunology (1993) 150:880-887, Clarkson et al., Nature (1991)
352:624-628).
[0122] Also encompassed by the present invention are polypeptides or
antibodies comprising variable chains having at least one conservative
amino acid substitution in at least one of the CDRs described herein (in
comparison with the original CDR).
[0123] The present invention also encompasses polypeptides or antibodies
comprising variable chains having at least one conservative amino acid
substitution in at least two of the CDRs (in comparison with the original
CDRs).
[0124] The present invention also encompasses polypeptides or antibodies
comprising variable chains having at least one conservative amino acid
substitution in the 3 CDRs (in comparison with the original CDRs).
[0125] The present invention also encompasses polypeptides or antibodies
comprising variable chains having at least two conservative amino acid
substitutions in at least one of the CDRs (in comparison with the
original CDRs).
[0126] The present invention also encompasses polypeptides or antibodies
comprising variable chains having at least two conservative amino acid
substitutions in at least two of the CDRs (in comparison with the
original CDRs).
[0127] The present invention also encompasses polypeptides or antibodies
comprising variable chains having at least two conservative amino acid
substitutions in the 3 CDRs (in comparison with the original CDRs).
[0128] In another aspect, the present invention relates to a polypeptide,
antibody or antigen binding fragment comprising (on a single polypeptide
chain or on separate polypeptide chains) at least one
complementarity-determining region of a light chain variable domain and
at least one complementarity-determining region of a heavy chain variable
domain of one of the antibodies or antigen binding fragment described
herein.
[0129] The present invention relates in another aspect thereof to
anti-KAAG1 antibodies that may comprise (on a single polypeptide chain or
on separate polypeptide chains) all six complementarity-determining
regions (CDRs) of the antibody or antigen binding fragment described
herein.
[0130] The antibodies or antigen binding fragment of the present invention
may further comprise additional amino acids flanking the amino and/or
carboxy region of the CDR(s). Those additional amino acids may be as
illustrated in Table A or Table B or may include, for example,
conservative amino acid substitution.
[0131] In accordance with the present invention, the antibody may comprise
a CDRL1 sequence comprising or consisting of formula:
X.sub.1aSSX.sub.2aSLLX.sub.3aX.sub.4aX.sub.5aX.sub.6aX.sub.7aX.sub.8aX.s-
ub.9aX.sub.10aLX.sub.11a (SEQ ID NO.:74)
wherein X.sub.1a may be a basic amino acid; wherein X.sub.2a may be a
basic amino acid; wherein X.sub.3a may be H, Y or N; wherein X.sub.4a may
be S, T, N or R; wherein X.sub.5a may be absent, S or N; wherein X.sub.6a
may be D, F or N; wherein X.sub.7a may be G or Q; wherein X.sub.8a may be
K, L or N; wherein X.sub.9a may be T or N; wherein X.sub.10a may be an
aromatic amino acid, and; wherein X.sub.11a may be A, N, E or Y.
[0132] In an exemplary embodiment of the invention X.sub.1a may be K or R.
[0133] In a further embodiment of the invention X.sub.2a may be Q or K.
[0134] In yet a further embodiment of the invention X.sub.3a may be N or
H.
[0135] In an additional embodiment of the invention X.sub.10a may be Y or
F.
[0136] More specific embodiments of the invention include CDRL1 of SEQ ID
NO.:74 where: X.sub.1a is K; X.sub.2a is Q; X.sub.3a is N; X.sub.3a is H;
X.sub.4a is S; X.sub.4a is T; X.sub.5a is S; X.sub.5a is absent; X.sub.6a
is N; X.sub.7a is Q; X.sub.7a is G; X.sub.8a is K; X.sub.9a is N;
X.sub.9a is T; X.sub.10a is Y; or X.sub.11a is A.
[0137] In accordance with the present invention, the antibody may comprise
a CDRL1 sequence comprising or consisting of formula:
KASQDX.sub.1bX.sub.2bX.sub.3bX.sub.4bX.sub.5bX.sub.6b (SEQ ID NO.:75)
wherein X.sub.1b may be an hydrophobic amino acid; wherein X.sub.2b may
be G or H; wherein X.sub.3b may be T, N or R; wherein X.sub.4b may be F,
Y or A; wherein X.sub.5b may be an hydrophobic amino acid, and; wherein
X.sub.6b may be N or A.
[0138] In an exemplary embodiment of the invention X.sub.1b may be V or I.
[0139] In another exemplary embodiment of the invention X.sub.5b may be V
or L.
[0140] More specific embodiments of the invention include CDRL1 of SEQ ID
NO.:75 where X.sub.1b is I; X.sub.2b is H; X.sub.3b is T; X.sub.3b is N;
X.sub.4b is Y; X.sub.4b is F; X.sub.5b is L or X.sub.6b is N.
[0141] In accordance with the present invention, the antibody may comprise
a CDRL2 sequence comprising or consisting of formula:
FX.sub.1cSTX.sub.2cX.sub.3cS (SEQ ID NO.:76)
Wherein X.sub.1c is A or G;
[0142] Wherein X.sub.2c is R or T, and;
Wherein X.sub.3c is E, K or A.
[0143] In an exemplary embodiment of the invention X.sub.1c may be A and
X.sub.2c may be T.
[0144] In another exemplary embodiment of the invention X.sub.1c may be A
and X.sub.2c may be R.
[0145] Other specific embodiments of the invention include CDRL2 of SEQ ID
NO.:76 where
X.sub.1c is A; X.sub.2c is R or X.sub.3c is E.
[0146] In accordance with the present invention, the antibody may comprise
a CDRL2 sequence comprising or consisting of formula:
X.sub.1dVSX.sub.2dX.sub.3dX.sub.4dS (SEQ ID NO.:77)
Wherein X.sub.1d may be L or K;
[0147] Wherein X.sub.2d may be a basic amino acid; Wherein X.sub.3d may be
L or R and;
Wherein X.sub.4d may be D or F.
[0148] In an exemplary embodiment of the invention X.sub.2d may be K or N.
[0149] Other specific embodiments of the invention include CDRL2 of SEQ ID
NO.:77 where X.sub.1d is L; X.sub.2d is K; X.sub.3d is L or X.sub.4d is
D.
[0150] In accordance with the present invention, the antibody may comprise
a CDRL2 sequence comprising or consisting of formula:
X.sub.1eANRLVX.sub.2e (SEQ ID NO.:78)
Wherein X.sub.1e may be a basic amino acid, and;
Wherein X.sub.2e may be D or A.
[0151] In an exemplary embodiment of the invention X.sub.1e may be R or H.
[0152] Other specific embodiments of the invention include CDRL2 of SEQ ID
NO.:78 where X.sub.1e is R or X.sub.2e is D.
[0153] In accordance with the present invention, the antibody may comprise
a CDRL3 sequence comprising or consisting of formula:
X.sub.1fQX.sub.2fX.sub.3fX.sub.4fX.sub.5fPLT (SEQ ID NO.:79)
Wherein X.sub.1f may be Q or L;
[0154] Wherein X.sub.2f may be an aromatic amino acid;
Wherein X.sub.3f may be D, F or Y;
[0155] Wherein X.sub.4f may be E, A, N or S, and;
Wherein X.sub.5f may be I, F or T.
[0156] In an exemplary embodiment of the invention X.sub.2f may be Y or H.
[0157] In another exemplary embodiment of the invention X.sub.3f may be Y
or D.
[0158] In yet another exemplary embodiment of the invention X.sub.5f may
be I or T.
[0159] Other specific embodiments of the invention include CDRL3 of SEQ ID
NO.:79 where X.sub.1f is Q; X.sub.2f is H; X.sub.3f is D; X.sub.3f is Y;
X.sub.4f is S; X.sub.4f is E; X.sub.4f is A; X.sub.5f is T, or X.sub.5f
is I.
[0160] In accordance with the present invention, the antibody may comprise
a CDRL3 sequence comprising or consisting of formula:
QQHX.sub.1gX.sub.2gX.sub.3gPLT (SEQ ID NO.:80)
Wherein X.sub.1g may be an aromatic amino acid; Wherein X.sub.2g may be N
or S, and;
Wherein X.sub.3g may be I or T.
[0161] In an exemplary embodiment of the invention X.sub.1g may be F or Y
[0162] Other specific embodiments of the invention include CDRL3 of SEQ ID
NO.:80 where X.sub.2g is S or X.sub.3g is T.
[0163] In accordance with the present invention, the antibody may comprise
a CDRL3 sequence comprising or consisting of formula:
X.sub.1hQGX.sub.2hHX.sub.3hPX.sub.4hT (SEQ ID NO.:81)
Wherein X.sub.1h may be an aromatic amino acid; Wherein X.sub.2h may be a
neutral hydrophilic amino acid; Wherein X.sub.3h may be F or V, and;
Wherein X.sub.4h may be R or L.
[0164] In an exemplary embodiment of the invention X.sub.1h may be W or F.
[0165] In another exemplary embodiment of the invention X.sub.2h may be S
or T.
[0166] Other specific embodiments of the invention include CDRL3 of SEQ ID
NO.:81 where X.sub.1h is W; X.sub.2h is T; X.sub.3h is F, or X.sub.4h is
R.
[0167] In accordance with the present invention, the antibody may comprise
a CDRH1 sequence comprising or consisting of formula:
GYX.sub.1iFX.sub.2iX.sub.3iYX.sub.4iX.sub.5iH (SEQ ID NO.:82)
Wherein X.sub.1i may be T, I or K;
[0168] Wherein X.sub.2i may be a neutral hydrophilic amino acid; Wherein
X.sub.3i may be an acidic amino acid; Wherein X.sub.4i may be E, N or D,
and; Wherein X.sub.5i may be hydrophobic amino acid.
[0169] In an exemplary embodiment of the invention X.sub.2i may be T or S.
[0170] In another exemplary embodiment of the invention X.sub.3i may be D
or E.
[0171] In yet another exemplary embodiment of the invention X.sub.4i may
be N or E.
[0172] In a further exemplary embodiment of the invention X.sub.5i may be
M, I or v.
[0173] Other specific embodiments of the invention include CDRH1 of SEQ ID
NO.:82 where X.sub.2i is T; X.sub.3i is D; X.sub.4i is E; X.sub.5i is I
or X.sub.5i is M.
[0174] In accordance with the present invention, the antibody may comprise
a CDRH2 sequence comprising or consisting of formula:
X.sub.1jX.sub.2jDPX.sub.3jTGX.sub.4jTX.sub.5j (SEQ ID NO.:83)
Wherein X.sub.1j may be V or G
[0175] Wherein X.sub.2j may be a hydrophobic amino acid;
Wherein X.sub.3j may be A, G or E;
[0176] Wherein X.sub.4j may be R, G, D, A, S, N or V, and; Wherein
X.sub.5j may be a hydrophobic amino acid.
[0177] In an exemplary embodiment of the invention X.sub.2j may be I or L.
[0178] In another exemplary embodiment of the invention X.sub.5j may be A
or V.
[0179] Other specific embodiments of the invention include CDRH2 of SEQ ID
NO.:83 where X.sub.1j is V; X.sub.2j is I; X.sub.3j is E; X.sub.4j is D
or X.sub.5j is A.
[0180] In accordance with the present invention, the antibody may comprise
a CDRH2 sequence comprising or consisting of formula:
VX.sub.1kDPX.sub.2kTGX.sub.3kTA (SEQ ID NO.:84)
Wherein X.sub.1k may be an hydrophobic amino acid;
Wherein X.sub.2k may be A, E or G;
Wherein X.sub.3k may be R, G, A, S, N V or D.
[0181] In an exemplary embodiment of the invention X.sub.1k may be L or I.
[0182] Other specific embodiments of the invention include CDRH2 of SEQ ID
NO.:84 where X.sub.1k is I; X.sub.2k is E, or X.sub.3k is D.
[0183] In accordance with the present invention, the antibody may comprise
a CDRH2 sequence comprising or consisting of formula:
YIX.sub.1lX.sub.2lX.sub.3lGX.sub.4lX.sub.5lX.sub.6l (SEQ ID NO.:85)
Wherein X.sub.1l may be S or N;
[0184] Wherein X.sub.2l may be an aromatic amino acid
Wherein X.sub.3l may be D, E or N;
Wherein X.sub.4l may be a D or H;
Wherein X.sub.5l may be Y, S or N;
Wherein X.sub.6l may be D, E or N.
[0185] In an exemplary embodiment of the invention X.sub.3l may be D or N.
[0186] In another exemplary embodiment of the invention X.sub.6l may be D
or N.
[0187] Other specific embodiments of the invention include CDRH2 of SEQ ID
NO.:85 where X.sub.2l is F or Y, X.sub.3l is N, X.sub.4l is D or X.sub.6l
is N.
[0188] In accordance with the present invention, the antibody may comprise
a CDRH2 sequence comprising or consisting of formula:
X.sub.1mINPYNX.sub.2mVTE (SEQ ID NO.:86)
wherein X.sub.1m may be N or Y, and; wherein X.sub.2m may be E, D or N.
[0189] In an exemplary embodiment of the invention X.sub.2m may be D or N.
[0190] Other specific embodiments of the invention include CDRH2 of SEQ ID
NO.:86 where
X.sub.1m is N or X.sub.2m is D.
[0191] In accordance with the present invention, the antibody may comprise
a CDRH2 sequence comprising or consisting of formula:
DINPX.sub.1nYGX.sub.2nX.sub.3nT (SEQ ID NO.:87)
Wherein X.sub.1n may be N or Y,
[0192] Wherein X.sub.2n may be G or T and; wherein X.sub.3n may be I or T.
[0193] In accordance with the present invention, the antibody may comprise
a CDRH3 sequence comprising or consisting of formula:
MX.sub.1oX.sub.2oX.sub.3oDY (SEQ ID NO.:88)
Wherein X.sub.1o may be G or S;
[0194] Wherein X.sub.2o may be Y or H, and; wherein X.sub.3o may be A or
S.
[0195] Other specific embodiments of the invention include CDRH3 of SEQ ID
NO.:88 where X.sub.1o is G; X.sub.2o is Y or X.sub.3o is S.
[0196] In accordance with the present invention, the antibody may comprise
a CDRH3 sequence comprising or consisting of formula:
IX.sub.1pYAX.sub.2pDY (SEQ ID NO.:89)
Wherein X.sub.1p may be G or S and; Wherein X.sub.2p may be absent or M.
[0197] Other specific embodiments of the invention include CDRH3 of SEQ ID
NO.:89 where X.sub.1p is S or X.sub.2p is M.
[0198] In accordance with the present invention, the antibody may comprise
a CDRH3 sequence comprising or consisting of formula:
AX.sub.1qX.sub.2qGLRX.sub.3q (SEQ ID NO.:90)
Wherein X.sub.1q may be R or W;
[0199] Wherein X.sub.2q may be an aromatic amino acid and; wherein
X.sub.3q may be a basic amino acid.
[0200] In an exemplary embodiment of the invention X.sub.2q may be W or F.
[0201] In another exemplary embodiment of the invention X.sub.3q may be Q
or N.
[0202] Other specific embodiments of the invention include CDRH3 of SEQ ID
NO.:90 where X.sub.1q is R; X.sub.2q is W or X.sub.3q is N.
[0203] The framework region of the heavy and/or light chains described
herein may be derived from one or more of the framework regions
illustrated in Tables A and B. The antibody or antigen binding fragments
may thus comprise one or more of the CDRs described herein (e.g.,
selected from the specific CDRs or consensus CDRs of SEQ ID NO.:74 to 90)
and framework regions originating from those illustrated in Tables A and
B. In Tables A and B, the expected CDRs are shown in bold, while the
framework regions are not.
[0204] Table 2 describes the sequences of the nucleotides and the amino
acids corresponding to the complete light and heavy chain immunoglobulins
of specific examples of anti-KAAG1 antibodies.
TABLE-US-00001
TABLE 2
complete sequences of light and heavy chain
immunoglobulins that bind to KAAG1
Nucleotide Amino acid
Antibody sequence sequence
designation Chain type (SEQ ID NO.:) (SEQ ID NO.:)
3D3 Light (L) 3 4
3D3 Heavy (H) 5 6
3G10 Light 7 8
3G10 Heavy 9 10
3C4 Light 11 12
3C4 Heavy 13 14
[0205] An antibody or antigen binding fragment that can bind KAAG1 may
comprise any one L chain with any one H chain immunoglobulin that is
listed in Table 2. In certain embodiments, the light chain of antibody
3D3 may be combined with the heavy chain of 3D3 or the heavy chain of
3G10 to form a complete antibody with KAAG1-binding activity. In an
exemplary embodiment of the present invention, the 3D3 L chain may be
combined with the 3D3 H chain, the 3G10 L chain may be combined with the
3G10 H chain, or the 3C4 L chain may be combined with the 3C4 H chain.
Additionally, some examples of antibodies or antigen binding fragment may
consist of any combination of two L chains and any two H chains from the
list of antibodies listed in Table 2.
[0206] The complete nucleotide sequences of the light and heavy
immunoglobulin chains of antibody 3D3 are shown in SEQ ID NOS:3 and 5,
respectively, and the corresponding amino acid sequences of the light and
heavy immunoglobulin chains of antibody 3D3 are shown in SEQ ID NOS:4 and
6, respectively. Thus, in an exemplary embodiment, an antibody that binds
to KAAG1 may comprise the light chain amino acid shown in SEQ ID NO.:4
combined with the heavy chain amino acid sequence shown in SEQ ID NO.:6.
In another embodiment, the antibody may comprise two identical 3D3 light
chains comprising of SEQ ID NO.:4 and two identical 3D3 heavy chains
comprising SEQ ID NO.:6.
[0207] The complete nucleotide sequences of the light and heavy
immunoglobulin chains of antibody 3G10 are shown in SEQ ID NOS:7 and 9,
respectively, and the corresponding amino acid sequences of the light and
heavy immunoglobulin chains of antibody 3G10 are shown in SEQ ID NOS:8
and 10, respectively. Thus, in an exemplary embodiment, an antibody that
binds to KAAG1 may comprise the light chain amino acid shown in SEQ ID
NO.:8 combined with the heavy chain amino acid sequence shown in SEQ ID
NO.:10. In another embodiment, the antibody may comprise two identical
3G10 light chains comprising SEQ ID NO.:8 and two identical 3G10 heavy
chains comprising SEQ ID NO.:10.
[0208] The complete nucleotide sequences of the light and heavy
immunoglobulin chains of antibody 3C4 are shown in SEQ ID NOS:11 and 13,
respectively and the corresponding amino acid sequences of the light and
heavy immunoglobulin chains of antibody 3C4 are shown in SEQ ID NOS:12
and 14, respectively. Thus, in an exemplary embodiment, an antibody that
binds to KAAG1 may comprise the light chain amino acid shown in SEQ ID
NO.:12 combined with the heavy chain amino acid sequence shown in SEQ ID
NO.:14. In another embodiment, the antibody may comprise two identical
3C4 light chains comprising SEQ ID NO.:12 and two identical 3C4 heavy
chains comprising SEQ ID NO.:14.
[0209] Variants of other anti-KAAG1 antibodies or antigen binding
fragments formed by the combination of light and/or heavy immunoglobulin
chains may each independently have at least 70%, 75%, 80%, 85%, 90%, 95%,
97%, or 99% identity to the amino acid sequences listed in Table 2 are
also provided. In certain embodiments, the antibody variants may comprise
at least one light chain and one heavy chain. In other instances, the
antibody variants may comprise two identical light chains and two
identical heavy chains. In accordance with the present invention, the
region of variation may be located in the constant region or in the
variable region. Also in accordance with the present invention, the
region of variation may be located in the framework region.
[0210] Also encompassed by the present invention are antibodies comprising
a light chain comprising one of the variable region illustrated in Table
A and a heavy chain comprising one of the variable region illustrated in
Table B. The light chain and heavy chain may comprise a constant domain.
Combinations of light chains and heavy chains of Table 2, Table A and
Table B are also encompassed by the present invention.
[0211] Antibodies or antigen binding fragments that contain the light
chain and heavy chain variable regions are also provided in the present
invention. Additionally, certain embodiments include antigen binding
fragments, variants, and derivatives of these light and heavy chain
variable regions.
[0212] Yet other exemplary embodiments of the invention includes an
isolated antibody or antigen binding fragment capable of specific binding
to SEQ ID NO.:2, to an extracellular portion of SEQ ID NO.:2, or to a
secreted form of SEQ ID NO.:2 or to a variant thereof, the antibody
comprising: [0213] a. the light chain variable domain defined in SEQ ID
NO.:16 and the heavy chain variable domain defined in SEQ ID NO.:18,
[0214] b. the light chain variable domain defined in SEQ ID NO.:20 and
the heavy chain variable domain defined in SEQ ID NO.:22; [0215] c. the
light chain variable domain defined in SEQ ID NO.:24 and the heavy chain
variable domain defined in SEQ ID NO.:26; [0216] d. the light chain
variable domain defined in SEQ ID NO.:105 and the heavy chain variable
domain defined in SEQ ID NO.:132, [0217] e. the light chain variable
domain defined in SEQ ID NO.:106 and the heavy chain variable domain
defined in SEQ ID NO.:133, [0218] f. the light chain variable domain
defined in SEQ ID NO.:107 and the heavy chain variable domain defined in
SEQ ID NO.:134, [0219] g. the light chain variable domain defined in SEQ
ID NO.:108 and the heavy chain variable domain defined in SEQ ID NO.:154,
[0220] h. the light chain variable domain defined in SEQ ID NO.:109 and
the heavy chain variable domain defined in SEQ ID NO.:153, [0221] i. the
light chain variable domain defined in SEQ ID NO.:110 and the heavy chain
variable domain defined in SEQ ID NO.:135, [0222] j. the light chain
variable domain defined in SEQ ID NO.:111 and the heavy chain variable
domain defined in SEQ ID NO.:136, [0223] k. the light chain variable
domain defined in SEQ ID NO.:112 and the heavy chain variable domain
defined in SEQ ID NO.:149, [0224] l. the light chain variable domain
defined in SEQ ID NO.:113 and the heavy chain variable domain defined in
SEQ ID NO.:137, [0225] m. the light chain variable domain defined in SEQ
ID NO.:114 and the heavy chain variable domain defined in SEQ ID NO.:140,
[0226] n. the light chain variable domain defined in SEQ ID NO.:115 and
the heavy chain variable domain defined in SEQ ID NO.:141, [0227] o. the
light chain variable domain defined in SEQ ID NO.:116 and the heavy chain
variable domain defined in SEQ ID NO.:142, [0228] p. the light chain
variable domain defined in SEQ ID NO.:117 and the heavy chain variable
domain defined in SEQ ID NO.:139, [0229] q. the light chain variable
domain defined in SEQ ID NO.:119 and the heavy chain variable domain
defined in SEQ ID NO.:143, [0230] r. the light chain variable domain
defined in SEQ ID NO.:120 and the heavy chain variable domain defined in
SEQ ID NO.:152, [0231] s. the light chain variable domain defined in SEQ
ID NO.:121 and the heavy chain variable domain defined in SEQ ID NO.:146,
[0232] t. the light chain variable domain defined in SEQ ID NO.:122 and
the heavy chain variable domain defined in SEQ ID NO.:138, [0233] u. the
light chain variable domain defined in SEQ ID NO.:123 and the heavy chain
variable domain defined in SEQ ID NO.:150, [0234] v. the light chain
variable domain defined in SEQ ID NO.:124 and the heavy chain variable
domain defined in SEQ ID NO.:144, [0235] w. the light chain variable
domain defined in SEQ ID NO.:126 and the heavy chain variable domain
defined in SEQ ID NO.:145, [0236] x. the light chain variable domain
defined in SEQ ID NO.:127 and the heavy chain variable domain defined in
SEQ ID NO.:157, [0237] y. the light chain variable domain defined in SEQ
ID NO.:128 and the heavy chain variable domain defined in SEQ ID NO.:155,
[0238] z. the light chain variable domain defined in SEQ ID NO.:129 and
the heavy chain variable domain defined in SEQ ID NO.:156, or; [0239] aa.
the light chain variable domain defined in SEQ ID NO.:130 and the heavy
chain variable domain defined in SEQ ID NO.:151.
[0240] It is to be understood herein, that the light chain variable region
of the specific combination provided above may be changed for any other
light chain variable region. Similarly, the heavy chain variable region
of the specific combination provided above may be changed for any other
heavy chain variable region.
[0241] Specific examples of sequences present in these light and heavy
chain variable regions are disclosed in Table 3.
TABLE-US-00002
TABLE 3
Sequences of light and heavy chain
variable regions that bind to KAAG1
Nucleotide Amino acid
Antibody Variable sequence sequence
designation region type (SEQ ID NO.:) (SEQ ID NO.:)
3D3 Light (VL) 15 16
3D3 Heavy (VH) 17 18
3G10 Light 19 20
3G10 Heavy 21 22
3C4 Light 23 24
3C4 Heavy 25 26
3z1A02 Light 105
3z1A02 Heavy 132
3z1E10 Light 109
3z1E10 Heavy 153
3z1G12L Light 126
3z1G12H Heavy 145
[0242] Therefore, antibodies and antigen binding fragments that bind to
KAAG1 may comprise one light variable region and one heavy variable
region of the same designated antibody or in any combinations. For
example, in an exemplary embodiment, an anti-KAAG1 antibody or fragment
may comprise the 3D3 light chain variable region (SEQ ID NO.:16) and the
3D3 heavy chain variable region (SEQ ID NO.:18). In an alternate
embodiment, an anti-KAAG1 antibody or fragment may comprise the 3D3 light
chain variable region (SEQ ID NO.:16) and the 3G10 heavy chain variable
region (SEQ ID NO.:22). In another embodiment, the anti-KAAG1 antibodies
may comprise two identical light chain variable regions and two identical
heavy chain regions. In yet another embodiment, the anti-KAAG1 antibodies
may comprise two different light chain variable regions and two different
heavy chain regions.
[0243] Variants of other anti-KAAG1 antibodies formed by the combination
of light and/or heavy chain variable regions that each have at least 70%,
75%, 80%, 85%, 90%, 95%, 97%, or 99% identity to the amino acid sequences
listed in Table 3 are also provided. Those skilled in the art will also
recognize that the anti-KAAG1 antibody variants may include conservative
amino acid changes, amino acid substitutions, deletions, or additions in
the amino acid sequences of the light and/or heavy chain variable regions
listed in Table 3.
[0244] In accordance with the present invention, the region of variation
may be located in the framework region of the variable region.
TABLE-US-00003
TABLE 4
Sequences of the light and heavy chain CDRs
Antibody Chain SEQ Amino acid
designation type CDR ID NO.: sequence
3D3 Light (L) CDR L1 27 KSSQSLLNSNFQKNFLA
3D3 Light CDR L2 28 FASTRES
3D3 Light CDR L3 29 QQHYSTPLT
3D3 Heavy (H) CDR H1 30 GYIFTDYEIH
3D3 Heavy CDR H2 31 VIDPETGNTA
3D3 Heavy CDR H3 32 MGYSDY
3G10 Light CDR L1 33 RSSQSLLHSNGNTYLE
3G10 Light CDR L2 34 KVSNRFS
3G10 Light CDR L3 35 FQGSHVPLT
3G10 Heavy CDR H1 36 GYTFTDNYMN
3G10 Heavy CDR H2 37 DINPYYGTTT
3G10 Heavy CDR H3 38 ARDDWFDY
3C4 Light CDR L1 39 KASQDIHNFLN
3C4 Light CDR L2 40 RANRLVD
3C4 Light CDR L3 41 LQYDEIPLT
3C4 Heavy CDR H1 42 GFSITSGYGWH
3C4 Heavy CDR H2 43 YINYDGHND
3C4 Heavy CDR H3 44 ASSYDGLFAY
3z1A02 Light CDR L1 158 KSSQSLLHSDGKTYLN
3z1A02 Light CDR L2 159 LVSKLDS
3z1A02 Light CDR L3 160 WQGTHFPRT
3z1A02 Heavy CDR H1 161 GYTFTD YNMH
3z1A02 Heavy CDR H2 162 YINPYNDVTE
3z1A02 Heavy CDR H3 163 AWFGL RQ
3z1E10 Light CDR L1 164 RSSKSLLHSNGN TYLY
3z1E10 Light CDR L2 165 RMSNLAS
3z1E10 Light CDR L3 166 MQHLEYPYT
3z1E10 Heavy CDR H1 167 GDTFTD YYMN
3z1E10 Heavy CDR H2 168 DINPNYGGIT
3z1E10 Heavy CDR H3 169 QAYYRNS DY
3z1G12L Light CDR L1 170 KASQDVGTAVA
3z1G12L Light CDR L2 171 WTSTRHT
3z1G12L Light CDR L3 172 QQHYSIPLT
3z1G12H Heavy CDR H1 173 GYIFTDYEIH
3z1G12H Heavy CDR H2 174 VIDPETGNTA
3z1G12H Heavy CDR H3 175 MGYSDY
[0245] In certain embodiments of the present invention, the anti-KAAG1
antibodies or antigen binding fragments may comprise the CDR sequences
shown in Table 4 or have substantial sequence identity to the CDR
sequences of Table 4. In an exemplary embodiment, the 3D3 anti-KAAG1
antibody may comprise a light chain variable region containing CDR1, 2,
and 3 that are encoded by SEQ ID NOS:27, 28, and 29, respectively, and/or
a heavy chain variable region containing CDR1, 2, and 3 that are encoded
by SEQ ID NOS:30, 31, and 32, respectively. In other embodiments the CDR3
region may be sufficient to provide antigen binding. As such polypeptides
comprising the CDR3 L or the CDR3H or both the CDR3 L and the CDR3H are
encompassed by the present invention.
[0246] Additionally, the anti-KAAG1 antibodies or antigen binding
fragments may include any combination of the CDRs listed in Table 4. For
example, the antibodies or antigen binding fragments may include the
light chain CDR3 and the heavy chain CDR3. It is understood that the CDRs
that are contained in the anti-KAAG1 antibodies or antigen binding
fragments may be variant CDRs with 80%, 85%, 90%, or 95% sequence
identity to the CDR sequences presented in Table 4. Those skilled in the
art will also recognize that the variants may include conservative amino
acid changes, amino acid substitutions, deletions, or additions in the
CDR sequences listed in Table 4.
[0247] Other exemplary embodiments of the invention includes an isolated
antibody or antigen binding fragment capable of specific binding to SEQ
ID NO.:2, to an extracellular portion of SEQ ID NO.:2 or to a secreted
form of SEQ ID NO.:2 or to a variant thereof, the antibody comprising:
[0248] a. the 3CDRs of a light chain variable domain defined in SEQ ID
NO.:16 and/or the 3CDRs of a heavy chain variable domain defined in SEQ
ID NO.:18, [0249] b. the 3CDRs of a light chain variable domain defined
in SEQ ID NO.:20 and/or the 3CDRs of a heavy chain variable domain
defined in SEQ ID NO.:22; [0250] c. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:24 and/or the 3CDRs of a heavy chain
variable domain defined in SEQ ID NO.:26; [0251] d. the 3CDRs of a light
chain variable domain defined in SEQ ID NO.:105 and/or the 3CDRs of a
heavy chain variable domain defined in SEQ ID NO.:132, [0252] e. the
3CDRs of a light chain variable domain defined in SEQ ID NO.:106 and/or
the 3CDRs of a heavy chain variable domain defined in SEQ ID NO.:133,
[0253] f. the 3CDRs of a light chain variable domain defined in SEQ ID
NO.:107 and/or the 3CDRs of a heavy chain variable domain defined in SEQ
ID NO.:134, [0254] g. the 3CDRs of a light chain variable domain defined
in SEQ ID NO.:108 and/or the 3CDRs of a heavy chain variable domain
defined in SEQ ID NO.:154, [0255] h. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:109 and/or the 3CDRs of a heavy chain
variable domain defined in SEQ ID NO.:153, [0256] i. the 3CDRs of a light
chain variable domain defined in SEQ ID NO.:110 and/or the 3CDRs of a
heavy chain variable domain defined in SEQ ID NO.:135, [0257] j. the
3CDRs of a light chain variable domain defined in SEQ ID NO.:111 and/or
the 3CDRs of a heavy chain variable domain defined in SEQ ID NO.:136,
[0258] k. the 3CDRs of a light chain variable domain defined in SEQ ID
NO.:112 and/or the 3CDRs of a heavy chain variable domain defined in SEQ
ID NO.:149, [0259] l. the 3CDRs of a light chain variable domain defined
in SEQ ID NO.:113 and/or the 3CDRs of a heavy chain variable domain
defined in SEQ ID NO.:137, [0260] m. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:114 and/or the 3CDRs of a heavy chain
variable domain defined in SEQ ID NO.:140, [0261] n. the 3CDRs of a light
chain variable domain defined in SEQ ID NO.:115 and/or the 3CDRs of a
heavy chain variable domain defined in SEQ ID NO.:141, [0262] o. the
3CDRs of a light chain variable domain defined in SEQ ID NO.:116 and/or
the 3CDRs of a heavy chain variable domain defined in SEQ ID NO.:142,
[0263] p. the 3CDRs of a light chain variable domain defined in SEQ ID
NO.:117 and/or the 3CDRs of a heavy chain variable domain defined in SEQ
ID NO.:139, [0264] q. the 3CDRs of a light chain variable domain defined
in SEQ ID NO.:119 and/or the 3CDRs of a heavy chain variable domain
defined in SEQ ID NO.:143, [0265] r. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:120 and/or the 3CDRs of a heavy chain
variable domain defined in SEQ ID NO.:152, [0266] s. the 3CDRs of a light
chain variable domain defined in SEQ ID NO.:121 and/or the 3CDRs of a
heavy chain variable domain defined in SEQ ID NO.:146, [0267] t. the
3CDRs of a light chain variable domain defined in SEQ ID NO.:122 and/or
the 3CDRs of a heavy chain variable domain defined in SEQ ID NO.:138,
[0268] u. the 3CDRs of a light chain variable domain defined in SEQ ID
NO.:123 and/or the 3CDRs of a heavy chain variable domain defined in SEQ
ID NO.:150, [0269] v. the 3CDRs of a light chain variable domain defined
in SEQ ID NO.:124 and/or the 3CDRs of a heavy chain variable domain
defined in SEQ ID NO.:144, [0270] w. the 3CDRs of a light chain variable
domain defined in SEQ ID NO.:126 and/or the 3CDRs of a heavy chain
variable domain defined in SEQ ID NO.:145, [0271] x. the 3CDRs of a light
chain variable domain defined in SEQ ID NO.:127 and/or the 3CDRs of a
heavy chain variable domain defined in SEQ ID NO.:157, [0272] y. the
3CDRs of a light chain variable domain defined in SEQ ID NO.:128 and/or
the 3CDRs of a heavy chain variable domain defined in SEQ ID NO.:155,
[0273] z. the 3CDRs of a light chain variable domain defined in SEQ ID
NO.:129 and/or the 3CDRs of a heavy chain variable domain defined in SEQ
ID NO.:156, or; [0274] aa. the 3CDRs of a light chain variable domain
defined in SEQ ID NO.:130 and/or the 3CDRs of a heavy chain variable
domain defined in SEQ ID NO.:151.
[0275] Again, the light chain variable region of the specific combination
provided above may be changed for any other light chain variable region
described herein. Similarly, the heavy chain variable region of the
specific combination provided above may be changed for any other heavy
chain variable region described herein.
Variant Antibody and Antigen Binding Fragments
[0276] The present invention also encompasses variants of the antibodies
or antigen binding fragments described herein. Variant antibodies or
antigen binding fragments included are those having a variation in the
amino acid sequence. For example, variant antibodies or antigen binding
fragments included are those having at least one variant CDR (two, three,
four, five or six variant CDRs or even twelve variant CDRs), a variant
light chain variable domain, a variant heavy chain variable domain, a
variant light chain and/or a variant heavy chain. Variant antibodies or
antigen binding fragments included in the present invention are those
having, for example, similar or improved binding affinity in comparison
with the original antibody or antigen binding fragment.
[0277] As used herein the term "variant" applies to any of the sequence
described herein and includes for example, a variant CDR (either CDRL1,
CDRL2, CDRL3, CDRH1, CDRH2 and/or CDRH3), a variant light chain variable
domain, a variant heavy chain variable domain, a variant light chain, a
variant heavy chain, a variant antibody, a variant antigen binding
fragment and a KAAG1 variant.
[0278] Variant antibodies or antigen binding fragments encompassed by the
present invention are those which may comprise an insertion, a deletion
or an amino acid substitution (conservative or non-conservative). These
variants may have at least one amino acid residue in its amino acid
sequence removed and a different residue inserted in its place.
[0279] The sites of greatest interest for substitutional mutagenesis
include the hypervariable regions (CDRs), but modifications in the
framework region or even in the constant region are also contemplated.
Conservative substitutions may be made by exchanging an amino acid (of a
CDR, variable chain, antibody, etc.) from one of the groups listed below
(group 1 to 6) for another amino acid of the same group.
[0280] Other exemplary embodiments of conservative substitutions are shown
in Table 1A under the heading of "preferred substitutions". If such
substitutions result in a undesired property, then more substantial
changes, denominated "exemplary substitutions" in Table 1A, or as further
described below in reference to amino acid classes, may be introduced and
the products screened.
[0281] It is known in the art that variants may be generated by
substitutional mutagenesis and retain the biological activity of the
polypeptides of the present invention. These variants have at least one
amino acid residue in the amino acid sequence removed and a different
residue inserted in its place. For example, one site of interest for
substitutional mutagenesis may include a site in which particular
residues obtained from various species are identical. Examples of
substitutions identified as "conservative substitutions" are shown in
Table 1A. If such substitutions result in a change not desired, then
other type of substitutions, denominated "exemplary substitutions" in
Table 1A, or as further described herein in reference to amino acid
classes, are introduced and the products screened.
[0282] Substantial modifications in function or immunological identity are
accomplished by selecting substitutions that differ significantly in
their effect on maintaining (a) the structure of the polypeptide backbone
in the area of the substitution, for example, as a sheet or helical
conformation. (b) the charge or hydrophobicity of the molecule at the
target site, or (c) the bulk of the side chain. Naturally occurring
residues are divided into groups based on common side chain properties:
[0283] (group 1) hydrophobic: norleucine, methionine (Met), Alanine
(Ala), Valine (Val), Leucine (Leu), Isoleucine (Ile) [0284] (group 2)
neutral hydrophilic: Cysteine (Cys), Serine (Ser), Threonine (Thr) [0285]
(group 3) acidic: Aspartic acid (Asp), Glutamic acid (Glu) [0286] (group
4) basic: Asparagine (Asn), Glutamine (Gin), Histidine (His), Lysine
(Lys), Arginine (Arg) [0287] (group 5) residues that influence chain
orientation: Glycine (Gly), Proline (Pro); and [0288] (group 6) aromatic:
Tryptophan (Trp), Tyrosine (Tyr), Phenylalanine (Phe)
[0289] Non-conservative substitutions will entail exchanging a member of
one of these classes for another.
TABLE-US-00004
TABLE 1A
Amino acid substitution
Original residue Exemplary substitution Conservative substitution
Ala (A) Val, Leu, Ile Val
Arg (R) Lys, Gln, Asn Lys
Asn (N) Gln, His, Lys, Arg, Asp Gln
Asp (D) Glu, Asn Glu
Cys (C) Ser, Ala Ser
Gln (Q) Asn; Glu Asn
Glu (E) Asp, Gln Asp
Gly (G) Ala Ala
His (H) Asn, Gln, Lys, Arg, Arg
Ile (I) Leu, Val, Met, Ala, Phe, Leu
norleucine
Leu (L) Norleucine, Ile, Val, Met, Ile
Ala, Phe
Lys (K) Arg, Gln, Asn Arg
Met (M) Leu, Phe, Ile Leu
Phe (F) Leu, Val, Ile, Ala, Tyr Tyr
Pro (P) Ala Ala
Ser (S) Thr Thr
Thr (T) Ser Ser
Trp (W) Tyr, Phe Tyr
Tyr (Y) Trp, Phe, Thr, Ser Phe
Val (V) Ile, Leu, Met, Phe, Ala, Leu
norleucine
[0290] Variation in the amino acid sequence of the variant antibody or
antigen binding fragment may include an amino acid addition, deletion,
insertion, substitution etc., one or more modification in the backbone or
side-chain of one or more amino acid, or an addition of a group or
another molecule to one or more amino acids (side-chains or backbone).
[0291] Variant antibody or antigen binding fragment may have substantial
sequence similarity and/or sequence identity in its amino acid sequence
in comparison with that the original antibody or antigen binding fragment
amino acid sequence. The degree of similarity between two sequences is
based upon the percentage of identities (identical amino acids) and of
conservative substitution.
[0292] Generally, the degree of similarity and identity between variable
chains has been determined herein using the Blast2 sequence program
(Tatiana A. Tatusova, Thomas L. Madden (1999), "Blast 2 sequences--a new
tool for comparing protein and nucleotide sequences", FEMS Microbiol
Lett. 174:247-250) using default settings, i.e., blastp program, BLOSUM62
matrix (open gap 11 and extension gap penalty 1; gapx dropoff 50, expect
10.0, word size 3) and activated filters.
[0293] Percent identity will therefore be indicative of amino acids which
are identical in comparison with the original peptide and which may
occupy the same or similar position.
[0294] Percent similarity will be indicative of amino acids which are
identical and those which are replaced with conservative amino acid
substitution in comparison with the original peptide at the same or
similar position.
[0295] Variants of the present invention therefore comprise those which
may have at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,
89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence
identity with an original sequence or a portion of an original sequence.
[0296] Exemplary embodiments of variants are those having at least 81%
sequence identity to a sequence described herein and 81%, 82%, 83%, 84%,
85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%
or 100% sequence similarity with an original sequence or a portion of an
original sequence.
[0297] Other exemplary embodiments of variants are those having at least
82% sequence identity to a sequence described herein and 82%, 83%, 84%,
85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%
or 100% sequence similarity with an original sequence or a portion of an
original sequence.
[0298] Further exemplary embodiments of variants are those having at least
85% sequence identity to a sequence described herein and 85%, 86%, 87%,
88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100%
sequence similarity with an original sequence or a portion of an original
sequence.
[0299] Other exemplary embodiments of variants are those having at least
90% sequence identity to a sequence described herein and 90%, 91%, 92%,
93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence similarity with an
original sequence or a portion of an original sequence.
[0300] Additional exemplary embodiments of variants are those having at
least 95% sequence identity to a sequence described herein and 95%, 96%,
97%, 98%, 99% or 100% sequence similarity with an original sequence or a
portion of an original sequence.
[0301] Yet additional exemplary embodiments of variants are those having
at least 97% sequence identity to a sequence described herein and 97%,
98%, 99% or 100% sequence similarity with an original sequence or a
portion of an original sequence.
[0302] For a purpose of concision the applicant provides herein a Table 1B
illustrating exemplary embodiments of individual variants encompassed by
the present invention and comprising the specified % sequence identity
and % sequence similarity. Each "X" is to be construed as defining a
given variant.
TABLE-US-00005
TABLE 1B
Percent (%) sequence identity
80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100
Percent (%) 80 X
sequence 81 X X
similarity 82 X X X
83 X X X X
84 X X X X X
85 X X X X X X
86 X X X X X X X
87 X X X X X X X X
88 X X X X X X X X X
89 X X X X X X X X X X
90 X X X X X X X X X X X
91 X X X X X X X X X X X X
92 X X X X X X X X X X X X X
93 X X X X X X X X X X X X X X
94 X X X X X X X X X X X X X X X
95 X X X X X X X X X X X X X X X X
96 X X X X X X X X X X X X X X X X X
97 X X X X X X X X X X X X X X X X X X
98 X X X X X X X X X X X X X X X X X X X
99 X X X X X X X X X X X X X X X X X X X X
100 X X X X X X X X X X X X X X X X X X X X X
[0303] The present invention encompasses CDRs, light chain variable
domains, heavy chain variable domains, light chains, heavy chains,
antibodies and/or antigen binding fragments which comprise at least 80%
identity with the sequence described herein.
[0304] Exemplary embodiments of the antibody or antigen binding fragment
of the present invention are those comprising a light chain variable
domain comprising a sequence selected from the group consisting of a
sequence at least 70%, 75%, 80% identical to SEQ ID NO.:16, a sequence at
least 70%, 75%, 80% identical to SEQ ID NO.:20, a sequence at least 70%,
75%, 80% identical to SEQ ID NO.:24, a sequence at least 70%, 75%, 80%
identical to SEQ ID NO.:105, a sequence at least 70%, 75%, 80% identical
to SEQ ID NO.:109 and a sequence at least 70%, 75%, 80% identical to SEQ
ID NO.:126.
[0305] These light chain variable domain may comprise a CDRL1 sequence at
least 80% identical to SEQ ID NO.:27, a CDRL2 sequence at least 80%
identical to SEQ ID NO.:28 and a CDRL3 sequence at least 80% identical to
SEQ ID NO.:29.
[0306] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRL1 sequence which may be at
least 90% identical to SEQ ID NO.:27.
[0307] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL1 sequence which may be
100% identical to SEQ ID NO.:27.
[0308] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence at least 90%
identical to SEQ ID NO.:28.
[0309] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL2 sequence which may
be 100% identical to SEQ ID NO.:28.
[0310] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
at least 90% identical to SEQ ID NO.:29.
[0311] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL3 sequence which may
be 100% identical to SEQ ID NO.:29.
[0312] The light chain variable domain listed above may comprise a CDRL1
sequence at least 80% identical to SEQ ID NO.:33, a CDRL2 sequence at
least 80% identical to SEQ ID NO.:34 and a CDRL3 sequence at least 80%
identical to SEQ ID NO.:35.
[0313] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRL1 sequence which may be at
least 90% identical to SEQ ID NO.:33.
[0314] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL1 sequence which may be
100% identical to SEQ ID NO.:33.
[0315] In yet a further exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL2 sequence which may
be at least 90% identical to SEQ ID NO.:34.
[0316] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence which may be
100% identical to SEQ ID NO.:34.
[0317] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
at least 90% identical to SEQ ID NO.:35.
[0318] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL3 sequence which may
be 100% identical to SEQ ID NO.:35.
[0319] The light chain variable domain listed above may comprise a CDRL1
sequence at least 80% identical to SEQ ID NO.:39, a CDRL2 sequence at
least 80% identical to SEQ ID NO.:40 and a CDRL3 sequence at least 80%
identical to SEQ ID NO.:41.
[0320] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRL1 sequence which may be at
least 90% identical to SEQ ID NO.:39.
[0321] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL1 sequence which may
be 100% identical to SEQ ID NO.:39.
[0322] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence which may be
at least 90% identical to SEQ ID NO.:40.
[0323] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence which may be
100% identical to SEQ ID NO.:40.
[0324] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL3 sequence which may
be at least 90% identical to SEQ ID NO.:41.
[0325] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
100% identical to SEQ ID NO.:41.
[0326] The light chain variable domain listed above may comprise a CDRL1
sequence at least 80% identical to SEQ ID NO.:158, a CDRL2 sequence at
least 80% identical to SEQ ID NO.:159 and a CDRL3 sequence at least 80%
identical to SEQ ID NO.:160.
[0327] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRL1 sequence which may be at
least 90% identical to SEQ ID NO.:158.
[0328] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL1 sequence which may be
100% identical to SEQ ID NO.:158.
[0329] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence which may be
at least 90% identical to SEQ ID NO.:159.
[0330] In yet a further exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL2 sequence which may
be 100% identical to SEQ ID NO.:159.
[0331] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
at least 90% identical to SEQ ID NO.:160.
[0332] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL3 sequence which may
be 100% identical to SEQ ID NO.:160.
[0333] The light chain variable domain listed above may comprise a CDRL1
sequence at least 80% identical to SEQ ID NO.:164, a CDRL2 sequence at
least 80% identical to SEQ ID NO.:165 and a CDRL3 sequence at least 80%
identical to SEQ ID NO.:166.
[0334] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRL1 sequence which may be at
least 90% identical to SEQ ID NO.:164.
[0335] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL1 sequence which may be
100% identical to SEQ ID NO.:164.
[0336] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL2 sequence which may
be at least 90% identical to SEQ ID NO.:165.
[0337] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence which may be
100% identical to SEQ ID NO.:165.
[0338] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL3 sequence which may
be at least 90% identical to SEQ ID NO.:166.
[0339] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
100% identical to SEQ ID NO.:166.
[0340] The light chain variable domain listed above may comprise a CDRL1
sequence at least 80% identical to SEQ ID NO.:170, a CDRL2 sequence at
least 80% identical to SEQ ID NO.:171 and a CDRL3 sequence at least 80%
identical to SEQ ID NO.:172.
[0341] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRL1 sequence which may be at
least 90% identical to SEQ ID NO.:170.
[0342] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL1 sequence which may
be 100% identical to SEQ ID NO.: 170.
[0343] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL2 sequence which may be
at least 90% identical to SEQ ID NO.: 171.
[0344] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRL2 sequence which may
be 100% identical to SEQ ID NO.: 171.
[0345] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
at least 90% identical to SEQ ID NO.: 172.
[0346] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRL3 sequence which may be
100% identical to SEQ ID NO.: 172.
[0347] An exemplary embodiment of a variant antibody light chain variable
region encompasses a light chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:16 and having up to 22 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:16. A SEQ ID NO.:16 variant is provided in SEQ ID NO.:178.
[0348] An exemplary embodiment of a variant antibody light chain variable
region encompasses a light chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:20 and having up to 22 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:20.
[0349] An exemplary embodiment of a variant antibody light chain variable
region encompasses a light chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:24 and having up to 21 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:24. A SEQ ID NO.:24 variant is provided in SEQ ID NO.:182.
[0350] An exemplary embodiment of a variant antibody light chain variable
region encompasses a light chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:105 and having up to 22 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:105.
[0351] An exemplary embodiment of a variant antibody light chain variable
region encompasses a light chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:109 and having up to 22 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:109.
[0352] An exemplary embodiment of a variant antibody light chain variable
region encompasses a light chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:126 and having up to 21 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:126.
[0353] In some instances, the variant antibody light chain variable region
may comprise amino acid deletions or additions (in combination or not
with amino acid substitutions). Often 1, 2, 3, 4 or 5 amino acid
deletions or additions may be tolerated.
[0354] In an exemplary embodiment, the antibody or antigen binding
fragment may comprise a heavy chain variable domain comprising a sequence
selected from the group consisting of a sequence at least 80% identical
to SEQ ID NO.:18, a sequence at least 70%, 75%, 80% identical to SEQ ID
NO.:22, a sequence at least 70%, 75%, 80% identical to SEQ ID NO.:26, a
sequence at least 70%, 75%, 80% identical to SEQ ID NO.:132, a sequence
at least 70%, 75%, 80% identical to SEQ ID NO.: 145 and a sequence at
least 70%, 75%, 80% identical to SEQ ID NO.:153.
[0355] These heavy chain variable domains may comprise a CDRH1 sequence at
least 80% identical to SEQ ID NO.:30, a CDRH2 sequence at least 80%
identical to SEQ ID NO.:31 and a CDRH3 sequence at least 80% identical to
SEQ ID NO.:32.
[0356] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH1 sequence which may be at
least 90% identical to SEQ ID NO.:30.
[0357] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH1 sequence which may be
100% identical to SEQ ID NO.:30.
[0358] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH2 sequence which may
be at least 90% identical to SEQ ID NO.:31.
[0359] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
100% identical to SEQ ID NO.:31.
[0360] In yet a further exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be at least 90% identical to SEQ ID NO.:32.
[0361] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be 100% identical to SEQ ID NO.:32.
[0362] The heavy chain variable domain listed above may comprise a CDRH1
sequence at least 80% identical to SEQ ID NO.:36, a CDRH2 sequence at
least 80% identical to SEQ ID NO.:37 and a CDRH3 sequence at least 80%
identical to SEQ ID NO.:38.
[0363] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH1 sequence which may be at
least 90% identical to SEQ ID NO.:36.
[0364] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH1 sequence which may be
100% identical to SEQ ID NO.:36.
[0365] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH2 sequence which may
be at least 90% identical to SEQ ID NO.:37.
[0366] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
100% identical to SEQ ID NO.:37.
[0367] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH3 sequence which may be
at least 90% identical to SEQ ID NO.:38.
[0368] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH3 sequence which may be
100% identical to SEQ ID NO.:38.
[0369] The heavy chain variable domain listed above may comprise a CDRH1
sequence at least 80% identical to SEQ ID NO.:42, a CDRH2 sequence at
least 80% identical to SEQ ID NO.:43 and a CDRH3 sequence at least 80%
identical to SEQ ID NO.:44.
[0370] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH1 sequence which may be
at least 90% identical to SEQ ID NO.:42.
[0371] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH1 sequence which may
be 100% identical to SEQ ID NO.:42.
[0372] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
at least 90% identical to SEQ ID NO.:43.
[0373] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
100% identical to SEQ ID NO.:43.
[0374] In yet a further exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be at least 90% identical to SEQ ID NO.:44.
[0375] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be 100% identical to SEQ ID NO.:44.
[0376] The heavy chain variable domain listed above may comprise a CDRH1
sequence at least 80% identical to SEQ ID NO.:161, a CDRH2 sequence at
least 80% identical to SEQ ID NO.:162 and a CDRH3 sequence at least 80%
identical to SEQ ID NO.:163.
[0377] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH1 sequence which may be at
least 90% identical to SEQ ID NO.:161.
[0378] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH1 sequence which may be
100% identical to SEQ ID NO.:161.
[0379] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
at least 90% identical to SEQ ID NO.:162.
[0380] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH2 sequence which may
be 100% identical to SEQ ID NO.:162.
[0381] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH3 sequence which may be
at least 90% identical to SEQ ID NO.:163.
[0382] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be 100% identical to SEQ ID NO.:163.
[0383] The heavy chain variable domain listed above may comprise a CDRH1
sequence at least 80% identical to SEQ ID NO.:167, a CDRH2 sequence at
least 80% identical to SEQ ID NO.:168 and a CDRH3 sequence at least 80%
identical to SEQ ID NO.:169.
[0384] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH1 sequence which may be at
least 90% identical to SEQ ID NO.:166.
[0385] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH1 sequence which may be
100% identical to SEQ ID NO.:166.
[0386] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH2 sequence which may
be at least 90% identical to SEQ ID NO.:168.
[0387] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
100% identical to SEQ ID NO.:168.
[0388] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be at least 90% identical to SEQ ID NO.:169.
[0389] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH3 sequence which may be
100% identical to SEQ ID NO.:169.
[0390] The heavy chain variable domain listed above may comprise a CDRH1
sequence at least 80% identical to SEQ ID NO.:173, a CDRH2 sequence at
least 80% identical to SEQ ID NO.:174 and a CDRH3 sequence at least 80%
identical to SEQ ID NO.:175.
[0391] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH1 sequence which may be at
least 90% identical to SEQ ID NO.:173.
[0392] In an exemplary embodiment of the present invention, any of the
antibodies provided herein may comprise a CDRH1 sequence which may be
100% identical to SEQ ID NO.: 173.
[0393] In an additional exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH2 sequence which may
be at least 90% identical to SEQ ID NO.: 174.
[0394] In a further exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH2 sequence which may be
100% identical to SEQ ID NO.: 174.
[0395] In another exemplary embodiment of the present invention, any of
the antibodies provided herein may comprise a CDRH3 sequence which may be
at least 90% identical to SEQ ID NO.: 175.
[0396] In yet another exemplary embodiment of the present invention, any
of the antibodies provided herein may comprise a CDRH3 sequence which may
be 100% identical to SEQ ID NO.: 175.
[0397] An exemplary embodiment of a variant antibody heavy chain variable
region encompasses a heavy chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:18 and having up to 22 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:18. A SEQ ID NO.:18 variant is provided in SEQ ID NO.:179.
[0398] An exemplary embodiment of a variant antibody heavy chain variable
region encompasses a heavy chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:22 and having up to 23 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:22.
[0399] An exemplary embodiment of a variant antibody heavy chain variable
region encompasses a heavy chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:26 and having up to 23 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:26. A SEQ ID NO.:26 variant is provided in SEQ ID NO.:183.
[0400] An exemplary embodiment of a variant antibody heavy chain variable
region encompasses a heavy chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:132 and having up to 23 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:132.
[0401] An exemplary embodiment of a variant antibody heavy chain variable
region encompasses a heavy chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:153 and having up to 23 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:153.
[0402] An exemplary embodiment of a variant antibody heavy chain variable
region encompasses a heavy chain variable region having CDR amino acid
sequences that are 100% identical to the CDR amino acid sequence of SEQ
ID NO.:145 and having up to 22 amino acid modifications (e.g.,
conservative or non-conservative amino acid substitutions) in its
framework region in comparison with the framework region of SEQ ID
NO.:145.
[0403] In some instances, the variant antibody heavy chain variable region
may comprise amino acid deletions or additions (in combination or not
with amino acid substitutions). Often 1, 2, 3, 4 or 5 amino acid
deletions or additions may be tolerated.
Production of the Antibodies in Cells
[0404] The anti-KAAG1 antibodies that are disclosed herein can be made by
a variety of methods familiar to those skilled in the art, such as
hybridoma methodology or by recombinant DNA methods.
[0405] In an exemplary embodiment of the invention, the anti-KAAG1
antibodies may be produced by the conventional hybridoma technology,
where a mouse is immunized with an antigen, spleen cells isolated and
fused with myeloma cells lacking HGPRT expression and hybrid cells
selected by hypoxanthine, aminopterin and thymine (HAT) containing media.
[0406] In an additional exemplary embodiment of the invention, the
anti-KAAG1 antibodies may be produced by recombinant DNA methods.
[0407] In order to express the anti-KAAG1 antibodies, nucleotide sequences
able to encode any one of a light and heavy immunoglobulin chains
described herein or any other may be inserted into an expression vector,
i.e., a vector that contains the elements for transcriptional and
translational control of the inserted coding sequence in a particular
host. These elements may include regulatory sequences, such as enhancers,
constitutive and inducible promoters, and 5' and 3' un-translated
regions. Methods that are well known to those skilled in the art may be
used to construct such expression vectors. These methods include in vitro
recombinant DNA techniques, synthetic techniques, and in vivo genetic
recombination.
[0408] A variety of expression vector/host cell systems known to those of
skill in the art may be utilized to express a polypeptide or RNA derived
from nucleotide sequences able to encode any one of a light and heavy
immunoglobulin chains described herein. These include, but are not
limited to, microorganisms such as bacteria transformed with recombinant
bacteriophage, plasmid, or cosmid DNA expression vectors; yeast
transformed with yeast expression vectors; insect cell systems infected
with baculovirus vectors; plant cell systems transformed with viral or
bacterial expression vectors; or animal cell systems. For long-term
production of recombinant proteins in mammalian systems, stable
expression in cell lines may be effected. For example, nucleotide
sequences able to encode any one of a light and heavy immunoglobulin
chains described herein may be transformed into cell lines using
expression vectors that may contain viral origins of replication and/or
endogenous expression elements and a selectable or visible marker gene on
the same or on a separate vector. The invention is not to be limited by
the vector or host cell employed. In certain embodiments of the present
invention, the nucleotide sequences able to encode any one of a light and
heavy immunoglobulin chains described herein may each be ligated into a
separate expression vector and each chain expressed separately. In
another embodiment, both the light and heavy chains able to encode any
one of a light and heavy immunoglobulin chains described herein may be
ligated into a single expression vector and expressed simultaneously.
[0409] Alternatively, RNA and/or polypeptide may be expressed from a
vector comprising nucleotide sequences able to encode any one of a light
and heavy immunoglobulin chains described herein using an in vitro
transcription system or a coupled in vitro transcription/translation
system respectively.
[0410] In general, host cells that contain nucleotide sequences able to
encode any one of a light and heavy immunoglobulin chains described
herein and/or that express a polypeptide encoded by the nucleotide
sequences able to encode any one of a light and heavy immunoglobulin
chains described herein, or a portion thereof, may be identified by a
variety of procedures known to those of skill in the art. These
procedures include, but are not limited to, DNA/DNA or DNA/RNA
hybridizations, PCR amplification, and protein bioassay or immunoassay
techniques that include membrane, solution, or chip based technologies
for the detection and/or quantification of nucleic acid or amino acid
sequences. Immunological methods for detecting and measuring the
expression of polypeptides using either specific polyclonal or monoclonal
antibodies are known in the art. Examples of such techniques include
enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs),
and fluorescence activated cell sorting (FACS). Those of skill in the art
may readily adapt these methodologies to the present invention.
[0411] Host cells comprising nucleotide sequences able to encode any one
of a light and heavy immunoglobulin chains described herein may thus be
cultured under conditions for the transcription of the corresponding RNA
(mRNA, siRNA, shRNA etc.) and/or the expression of the polypeptide from
cell culture. The polypeptide produced by a cell may be secreted or may
be retained intracellularly depending on the sequence and/or the vector
used. In an exemplary embodiment, expression vectors containing
nucleotide sequences able to encode any one of a light and heavy
immunoglobulin chains described herein may be designed to contain signal
sequences that direct secretion of the polypeptide through a prokaryotic
or eukaryotic cell membrane.
[0412] Due to the inherent degeneracy of the genetic code, other DNA
sequences that encode the same, substantially the same or a functionally
equivalent amino acid sequence may be produced and used, for example, to
express a polypeptide encoded by nucleotide sequences able to encode any
one of a light and heavy immunoglobulin chains described herein. The
nucleotide sequences of the present invention may be engineered using
methods generally known in the art in order to alter the nucleotide
sequences for a variety of purposes including, but not limited to,
modification of the cloning, processing, and/or expression of the gene
product. DNA shuffling by random fragmentation and PCR reassembly of gene
fragments and synthetic oligonucleotides may be used to engineer the
nucleotide sequences. For example, oligonucleotide-mediated site-directed
mutagenesis may be used to introduce mutations that create new
restriction sites, alter glycosylation patterns, change codon preference,
produce splice variants, and so forth.
[0413] In addition, a host cell strain may be chosen for its ability to
modulate expression of the inserted sequences or to process the expressed
polypeptide in the desired fashion. Such modifications of the polypeptide
include, but are not limited to, acetylation, carboxylation,
glycosylation, phosphorylation, lipidation, and acylation. In an
exemplary embodiment, anti-KAAG1 antibodies that contain particular
glycosylation structures or patterns may be desired. Post-translational
processing, which cleaves a "prepro" form of the polypeptide, may also be
used to specify protein targeting, folding, and/or activity. Different
host cells that have specific cellular machinery and characteristic
mechanisms for post-translational activities (e.g., CHO, HeLa, MDCK,
HEK293, and W138) are available commercially and from the American Type
Culture Collection (ATCC) and may be chosen to ensure the correct
modification and processing of the expressed polypeptide.
[0414] Those of skill in the art will readily appreciate that natural,
modified, or recombinant nucleic acid sequences may be ligated to a
heterologous sequence resulting in translation of a fusion polypeptide
containing heterologous polypeptide moieties in any of the aforementioned
host systems. Such heterologous polypeptide moieties may facilitate
purification of fusion polypeptides using commercially available affinity
matrices. Such moieties include, but are not limited to, glutathione
S-transferase (GST), maltose binding protein, thioredoxin, calmodulin
binding peptide, 6-His (His), FLAG, c-myc, hemaglutinin (HA), and
antibody epitopes such as monoclonal antibody epitopes.
[0415] In yet a further aspect, the present invention relates to a
polynucleotide which may comprise a nucleotide sequence encoding a fusion
protein. The fusion protein may comprise a fusion partner (e.g., HA, Fc,
etc.) fused to the polypeptide (e.g., complete light chain, complete
heavy chain, variable regions, CDRs etc.) described herein.
[0416] Those of skill in the art will also readily recognize that the
nucleic acid and polypeptide sequences may be synthesized, in whole or in
part, using chemical or enzymatic methods well known in the art. For
example, peptide synthesis may be performed using various solid-phase
techniques and machines such as the ABI 431A Peptide synthesizer (PE
Biosystems) may be used to automate synthesis. If desired, the amino acid
sequence may be altered during synthesis and/or combined with sequences
from other proteins to produce a variant protein.
Antibody Conjugates
[0417] The antibody or antigen binding fragment of the present invention
may be conjugated with a detectable moiety (i.e., for detection or
diagnostic purposes) or with a therapeutic moiety (for therapeutic
purposes)
[0418] A "detectable moiety" is a moiety detectable by spectroscopic,
photochemical, biochemical, immunochemical, chemical and/or other
physical means. A detectable moiety may be coupled either directly and/or
indirectly (for example via a linkage, such as, without limitation, a
DOTA or NHS linkage) to antibodies and antigen binding fragments thereof
of the present invention using methods well known in the art. A wide
variety of detectable moieties may be used, with the choice depending on
the sensitivity required, ease of conjugation, stability requirements and
available instrumentation. A suitable detectable moiety include, but is
not limited to, a fluorescent label, a radioactive label (for example,
without limitation, .sup.125I, In.sup.111, Tc.sup.99, I.sup.131 and
including positron emitting isotopes for PET scanner etc), a nuclear
magnetic resonance active label, a luminiscent label, a chemiluminescent
label, a chromophore label, an enzyme label (for example and without
limitation horseradish peroxidase, alkaline phosphatase, etc.), quantum
dots and/or a nanoparticle. Detectable moiety may cause and/or produce a
detectable signal thereby allowing for a signal from the detectable
moiety to be detected.
[0419] In another exemplary embodiment of the invention, the antibody or
antigen binding fragment thereof may be coupled (modified) with a
therapeutic moiety (e.g., drug, cytotoxic moiety).
[0420] In an exemplary embodiment, the anti-KAAG1 antibodies and antigen
binding fragments may comprise a chemotherapeutic or cytotoxic agent. For
example, the antibody and antigen binding fragments may be conjugated to
the chemotherapeutic or cytotoxic agent. Such chemotherapeutic or
cytotoxic agents include, but are not limited to, Yttrium-90,
Scandium-47, Rhenium-186, Iodine-131, Iodine-125, and many others
recognized by those skilled in the art (e.g., lutetium (e.g.,
Lu.sup.177), bismuth (e.g., Bi.sup.213), copper (e.g., Cu.sup.67)). In
other instances, the chemotherapeutic or cytotoxic agent may be comprised
of, among others known to those skilled in the art, 5-fluorouracil,
adriamycin, irinotecan, taxanes, pseudomonas endotoxin, ricin and other
toxins.
[0421] Alternatively, in order to carry out the methods of the present
invention and as known in the art, the antibody or antigen binding
fragment of the present invention (conjugated or not) may be used in
combination with a second molecule (e.g., a secondary antibody, etc.)
which is able to specifically bind to the antibody or antigen binding
fragment of the present invention and which may carry a desirable
detectable, diagnostic or therapeutic moiety.
Pharmaceutical Compositions of the Antibodies and their Use
[0422] Pharmaceutical compositions of the anti-KAAG1 antibodies
(conjugated or not) are also encompassed by the present invention. The
pharmaceutical composition may comprise an anti-KAAG1 antibody or an
antigen binding fragment and may also contain a pharmaceutically
acceptable carrier.
[0423] Other aspects of the invention relate to a composition which may
comprise the antibody or antigen binding fragment described herein and a
carrier.
[0424] The present invention also relates to a pharmaceutical composition
which may comprise the antibody or antigen binding fragment described
herein and a pharmaceutically acceptable carrier.
[0425] Yet other aspects of the invention relate to the use of the
isolated antibody or antigen binding fragment described herein in the
treatment or diagnosis of ovarian cancer.
[0426] In addition to the active ingredients, a pharmaceutical composition
may contain pharmaceutically acceptable carriers comprising water, PBS,
salt solutions, gelatins, oils, alcohols, and other excipients and
auxiliaries that facilitate processing of the active compounds into
preparations that may be used pharmaceutically. In other instances, such
preparations may be sterilized.
[0427] As used herein, "pharmaceutical composition" means therapeutically
effective amounts of the agent together with pharmaceutically acceptable
diluents, preservatives, solubilizers, emulsifiers, adjuvant and/or
carriers. A "therapeutically effective amount" as used herein refers to
that amount which provides a therapeutic effect for a given condition and
administration regimen. Such compositions are liquids or lyophilized or
otherwise dried formulations and include diluents of various buffer
content (e.g., Tris-HCl, acetate, phosphate), pH and ionic strength,
additives such as albumin or gelatin to prevent absorption to surfaces,
detergents (e.g., polysorbate 20 (TWEEN.TM. 20), polysorbate 80
(TWEEN.TM. 80), the polyoxyethylene-polyoxypropylene block copolymer:
PLURONIC.TM. F68, bile acid salts). Solubilizing agents (e.g., glycerol,
polyethylene glycerol), anti-oxidants (e.g., ascorbic acid, sodium
metabisulfite), preservatives (e.g., thimerosal, benzyl alcohol,
parabens), bulking substances or tonicity modifiers (e.g., lactose,
mannitol), covalent attachment of polymers such as polyethylene glycol to
the protein, complexation with metal ions, or incorporation of the
material into or onto particulate preparations of polymeric compounds
such as polylactic acid, polyglycolic acid, hydrogels, etc, or onto
liposomes, microemulsions, micelles, unilamellar or multilamellar
vesicles, erythrocyte ghosts, or spheroplasts. Such compositions will
influence the physical state, solubility, stability, rate of in vivo
release, and rate of in vivo clearance. Controlled or sustained release
compositions include formulation in lipophilic depots (e.g., fatty acids,
waxes, oils). Also comprehended by the invention are particulate
compositions coated with polymers (e.g., poloxamers or poloxamines).
Other embodiments of the compositions of the invention incorporate
particulate forms protective coatings, protease inhibitors or permeation
enhancers for various routes of administration, including parenteral,
pulmonary, nasal, oral, vaginal, rectal routes. In one embodiment the
pharmaceutical composition is administered parenterally, paracancerally,
transmucosally, transdermally, intramuscularly, intravenously,
intradermally, subcutaneously, intraperitonealy, intraventricularly,
intracranially and intratumorally.
[0428] Further, as used herein "pharmaceutically acceptable carrier" or
"pharmaceutical carrier" are known in the art and include, but are not
limited to, 0.01-0.1 M or 0.05 M phosphate buffer or 0.8% saline.
Additionally, such pharmaceutically acceptable carriers may be aqueous or
non-aqueous solutions, suspensions, and emulsions. Examples of
non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable
oils such as olive oil, and injectable organic esters such as ethyl
oleate. Aqueous carriers include water, alcoholic/aqueous solutions,
emulsions or suspensions, including saline and buffered media. Parenteral
vehicles include sodium chloride solution, Ringer's dextrose, dextrose
and sodium chloride, lactated Ringer's orfixed oils. Intravenous vehicles
include fluid and nutrient replenishers, electrolyte replenishers such as
those based on Ringer's dextrose, and the like. Preservatives and other
additives may also be present, such as, for example, antimicrobials,
antioxidants, collating agents, inert gases and the like.
[0429] For any compound, the therapeutically effective dose may be
estimated initially either in cell culture assays or in animal models
such as mice, rats, rabbits, dogs, or pigs. An animal model may also be
used to determine the concentration range and route of administration.
Such information may then be used to determine useful doses and routes
for administration in humans. These techniques are well known to one
skilled in the art and a therapeutically effective dose refers to that
amount of active ingredient that ameliorates the symptoms or condition.
Therapeutic efficacy and toxicity may be determined by standard
pharmaceutical procedures in cell cultures or with experimental animals,
such as by calculating and contrasting the ED.sub.50 (the dose
therapeutically effective in 50% of the population) and LD.sub.50 (the
dose lethal to 50% of the population) statistics. Any of the therapeutic
compositions described above may be applied to any subject in need of
such therapy, including, but not limited to, mammals such as dogs, cats,
cows, horses, rabbits, monkeys, and humans.
[0430] The pharmaceutical compositions utilized in this invention may be
administered by any number of routes including, but not limited to, oral,
intravenous, intramuscular, intra-arterial, intramedullary, intrathecal,
intraventricular, transdermal, subcutaneous, intraperitoneal, intranasal,
enteral, topical, sublingual, or rectal means.
[0431] The term "treatment" for purposes of this disclosure refers to both
therapeutic treatment and prophylactic or preventative measures, wherein
the object is to prevent or slow down (lessen) the targeted pathologic
condition or disorder. Those in need of treatment include those already
with the disorder as well as those prone to have the disorder or those in
whom the disorder is to be prevented.
[0432] The anti-KAAG1 antibodies and antigen binding fragments therein may
have therapeutic uses in the treatment of various cancer types, such as
ovarian cancer, renal cancer, colon cancer, lung cancer, melanoma, etc.
In an exemplary embodiment, the antibodies and fragments have therapeutic
uses in ovarian cancer. In certain instances, the anti-KAAG1 antibodies
and fragments may interact with cancer cells that express KAAG1 and
induce an immunological reaction by mediating ADCC. In other instances,
the anti-KAAG1 antibodies and fragments may block the interaction of
KAAG1 with its protein partners.
[0433] The anti-KAAG1 antibodies and antigen binding fragments therein may
have therapeutic uses in the treatment of various types of ovarian
cancer. Several different cell types may give rise to different ovarian
cancer histotypes. The most common form of ovarian cancer is comprised of
tumors that originate in the epithelial cell layer of the ovary or the
fallopian tube. Such epithelial ovarian cancers include serous tumors,
endometroid tumors, mucinous tumors, clear cell tumors, and borderline
tumors. In other embodiments, the anti-KAAG1 antibodies and antigen
binding fragments therein have uses in the treatment of other types of
ovarian cancer such as germ line and sex cord ovarian cancer.
[0434] In certain instances, the anti-KAAG1 antibodies and antigen binding
fragments therein may be administered concurrently in combination with
other treatments given for the same condition. As such, the antibodies
may be administered with anti-mitotics (eg., taxanes), platinum-based
agents (eg., cisplatin), DNA damaging agents (eg. Doxorubicin) and other
anti-cancer therapies that are known to those skilled in the art. In
other instances, the anti-KAAG1 antibodies and antigen binding fragments
therein may be administered with other therapeutic antibodies. These
include, but are not limited to, antibodies that target EGFR, CD-20, and
Her2.
[0435] The present invention relates in a further aspect thereof to a
method for inhibiting the growth of a KAAG1-expressing cell, the method
which may comprise contacting the cell with an effective amount of the
antibody or antigen binding fragment described herein.
[0436] The present invention also encompasses method of treating cancer or
inhibiting the growth of a KAAG1 expressing cells in a mammal, the method
may comprise administering the antibody or antigen binding fragment
described herein to a mammal in need.
[0437] In further aspects, the present invention provides method of
treatment, diagnostic methods and method of detection using the antibody
or antigen binding fragment of the present invention and the use of these
antibodies or antigen binding fragment in the manufacture of a
pharmaceutical composition or drug for such purposes.
[0438] Method of treatment encompassed by the present invention includes
administering an antibody or antigen binding fragment described herein to
a mammal in need, and especially to a patient having or susceptible of
having a cancer.
[0439] The invention also provides in further aspects, methods for
reducing tumor spread, tumor invasion, tumor formation or for inducing
tumor lysis, which may comprise administering an isolated antibody or
antigen binding fragment to a mammal in need.
[0440] The invention therefore relates to the use of the isolated antibody
described herein in the (manufacture of a pharmaceutical composition for)
treatment of cancer, reduction of tumor spread, tumor invasion, tumor
formation or for inducing tumor lysis of KAAG1-expressing tumor cells.
[0441] The antibody or antigen binding fragment may more particularly be
applicable for malignant tumor including, for example, a malignant tumor
having the ability to metastasize and/or tumor cells characterized by
anchorage-independent growth. The antibody or antigen binding fragment of
the present invention may also be used in the diagnosis of cancer. The
diagnosis of cancer may be performed in vivo by administering the
antibody or antigen binding fragment of the present invention to a mammal
having or suspected of having a cancer. The diagnosis may also be
performed ex vivo by contacting a sample obtained from the mammal with
the antibody or antigen binding fragment and determining the presence or
absence of cells (tumor cells) expressing KAAG1.
[0442] The present invention also encompasses method of detecting cancer
or detecting a KAAG1 expressing cells in a mammal, the method may
comprise administering the antibody or antigen binding fragment described
herein to a mammal in need.
[0443] The present invention relates in another aspect thereof to a method
for detecting a KAAG1-expressing cell, the method may comprise contacting
the cell with an antibody or antigen binding fragment described herein
and detecting a complex formed by the antibody and the KAAG1-expressing
cell. Exemplary embodiments of antibodies or antigen binding fragments
used in detection methods are those which are capable of binding to the
extracellular region of KAAG1.
[0444] Other exemplary embodiments of antibodies or antigen binding
fragments used in detection methods are those which bind to KAAG1
expressed at the surface of a tumor cells.
[0445] Patients which would benefit from treatment, detection or
diagnostic methods described herein are those which have or are suspected
of having ovarian cancer (e.g., serous, endometroid, clear cell or
mucinous), skin cancer (e.g., melanomas, squamous cell carcinomas), renal
cancer (e.g., papillary cell carcinomas, clear cell carcinomas),
colorectal cancer (e.g., colorectal carcinomas), sarcoma, leukemia, brain
tumor, thyroid tumor, breast cancer (e.g., mammary carcinomas), prostate
cancer (e.g., prostatic carcinomas), oesophageal tumor, bladder tumor,
lung tumor (e.g., lung carcinomas) or head and neck tumor and especially
when the cancer is characterized as being malignant and/or when the
KAAG1-expressing cells are characterized by anchorage-independent growth.
[0446] Especially encompassed by the present invention are patients having
or susceptible of having ovarian cancer (e.g., serous, endometroid, clear
cell or mucinous), skin cancer (e.g., melanomas, squamous cell
carcinomas) or renal cancer (e.g., papillary cell carcinomas) and
especially when the cancer is characterized as being malignant and/or
when the KAAG1-expressing cells are characterized by
anchorage-independent growth.
[0447] Another aspect of the invention relates a method for detecting
KAAG1 (SEQ ID NO.:2), a KAAG1 variant having at least 80% sequence
identity with SEQ ID NO.:2 or a secreted form of circulating form of
KAAG1 or KAAG1 variant, the method may comprise contacting a cell
expressing KAAG1 or the KAAG1 variant or a sample (biopsy, serum, plasma,
urine etc.) comprising or suspected of comprising KAAG1 or the KAAG1
variant with the antibody or antigen binding fragments described herein
and measuring binding. The sample may originate from a mammal (e.g., a
human) which may have cancer (e.g., ovarian cancer) or may be suspected
of having cancer (e.g., ovarian cancer). The sample may be a tissue
sample obtained from the mammal or a cell culture supernatant.
[0448] In accordance with the invention the sample may be a serum sample,
a plasma sample, a blood sample or ascitic fluid obtained from the
mammal. The antibody or antigen binding fragment described herein may
advantageously detect a secreted or circulating form (circulating in
blood) of KAAG1.
[0449] The method may comprise quantifying the complex formed by the
antibody or antigen binding fragment bound to KAAG1 or to the KAAG1
variant.
[0450] The binding of an antibody to an antigen will cause an increase in
the expected molecular weight of the antigen. A physical change therefore
occurs upon specific binding of the antibody or antigen binding fragment
and the antigen.
[0451] Such changes may be detected using, for example, electrophoresis
followed by Western blot and coloration of the gel or blot, mass
spectrometry, HPLC coupled with a computer or else. Apparatus capable of
computing a shift in molecular weight are known in the art and include
for example, PHOSPHORIMAGER.TM..
[0452] When the antibody comprises for example a detectable label, the
antigen-antibody complex may be detected by the fluorescence emitted by
the label, radiation emission of the label, enzymatic activity of a label
provided with its substrate or else.
[0453] Detection and/or measurement of binding between an antibody or
antigen binding fragment and an antigen may be performed by various
methods known in the art. Binding between an antibody or antigen binding
fragment and an antigen may be monitored with an apparatus capable of
detecting the signal emitted by the detectable label (radiation emission,
fluorescence, color change etc.). Such apparatus provides data which
indicates that binding as occurred and may also provide indication as to
the amount of antibody bound to the antigen. The apparatus (usually
coupled with a computer) may also be capable of calculating the
difference between a background signal (e.g., signal obtained in the
absence of antigen-antibody binding) or background noise and the signal
obtained upon specific antibody-antigen binding. Such apparatuses may
thus provide the user with indications and conclusions as to whether the
antigen has been detected or not.
[0454] Additional aspects of the invention relates to kits which may
include one or more container containing one or more antibodies or
antigen binding fragments described herein.
Nucleic Acids, Vectors and Cells
[0455] Antibodies are usually made in cells allowing expression of the
light chain and heavy chain expressed from a vector(s) comprising a
nucleic acid sequence encoding the light chain and heavy chain.
[0456] The present therefore encompasses nucleic acids capable of encoding
any of the CDRs, light chain variable domains, heavy chain variable
domains, light chains, heavy chains described herein.
[0457] The present invention therefore relates in a further aspect to a
nucleic acid encoding a light chain variable domain and/or a heavy chain
variable domain of an antibody which is capable of specific binding to
KAAG1.
[0458] In accordance with an embodiment of the invention, the nucleic acid
may especially encode a light chain variable domain and/or heavy chain
variable domain of an antibody which may be capable of inducing killing
(elimination, destruction, lysis) of KAAG1-expressing tumor cells.
[0459] In accordance with another embodiment of the invention, the nucleic
acid may especially encode a light chain variable domain and/or heavy
chain variable domain of an antibody which may be capable of reducing
spreading of KAAG1-expressing tumor cells.
[0460] In accordance with yet another embodiment of the invention, the
nucleic acid may particularly encode a light chain variable domain and/or
heavy chain variable domain of an antibody which may be capable of
decreasing or impairing formation of KAAG1-expressing tumors.
[0461] Exemplary embodiments of nucleic acids of the present invention
include nucleic acids encoding a light chain variable domain comprising:
[0462] a. a CDRL1 sequence selected from the group consisting of SEQ ID
NO.:74 and SEQ ID NO.:75; [0463] b. a CDRL2 sequence selected from the
group consisting of SEQ ID NO.:76, SEQ ID NO.: 77 and SEQ ID NO.:78, or;
[0464] c. a CDRL3 sequence selected from the group consisting of SEQ ID
NO.:79, SEQ ID NO.:80 and SEQ ID NO.:81.
[0465] In accordance with the present invention, the nucleic acid may
encode a light chain variable domain which may comprise at least two CDRs
of a CDRL1, a CDRL2 or a CDRL3.
[0466] Also in accordance with the present invention, the nucleic acid may
encode a light chain variable domain which may comprise one CDRL1, one
CDRL2 and one CDRL3.
[0467] The present invention also relates to a nucleic acid encoding a
heavy chain variable domain comprising: [0468] a. a CDRH1 sequence
comprising SEQ ID NO.:82; [0469] b. a CDRH2 sequence selected from the
group consisting of SEQ ID NO.:83, SEQ ID NO.:84, SEQ ID NO.:85, SEQ ID
NO.:86 and SEQ ID NO.:87, or; [0470] c. a CDRH3 sequence selected from
the group consisting of SEQ ID NO.:88, SEQ ID NO.:89 and SEQ ID NO.:90.
[0471] In accordance with the present invention, the nucleic acid may
encode a heavy chain variable domain which may comprise at least two CDRs
of a CDRH1, a CDRH2 or a CDRH3.
[0472] In accordance with the present invention, the nucleic acid may
encode a heavy chain variable domain which may comprise one CDRH1, one
CDRH2 and one CDRH3.
[0473] Also encompassed by the present invention are nucleic acids
encoding antibody variants having at least one conservative amino acid
substitution.
[0474] In accordance with the present invention, the nucleic acid may
encode a CDR comprising at least one conservative amino acid
substitution.
[0475] In accordance with the present invention, the nucleic acid may
encode a CDR comprising at least one conservative amino acid substitution
in at least two of the CDRs.
[0476] In accordance with the present invention, the nucleic acid may
encode a CDR comprising at least one conservative amino acid substitution
in the 3 CDRs.
[0477] In accordance with the present invention, the nucleic acid may
encode a CDR comprising at least two conservative amino acid
substitutions in at least one of the CDRs.
[0478] In accordance with the present invention, the nucleic acid may
encode a CDR comprising at least two conservative amino acid
substitutions in at least two of the CDRs.
[0479] In accordance with the present invention, the nucleic acid may
encode a CDR comprising at least two conservative amino acid
substitutions in the 3 CDRs.
[0480] Other aspects of the invention relate to a nucleic acid encoding a
light chain variable domain having at least 70%, 75%, 80% sequence
identity to a sequence selected from the group consisting of SEQ ID
NO.:16, SEQ ID NO.:20, SEQ ID NO.:24, SEQ ID NO.:105, SEQ ID NO.:106, SEQ
ID NO.:107, SEQ ID NO.:108, SEQ ID NO.:109, SEQ ID NO.:110, SEQ ID
NO.:111, SEQ ID NO.:112, SEQ ID NO.:113, SEQ ID NO.:114, SEQ ID NO.:115,
SEQ ID NO.:116, SEQ ID NO.:117, SEQ ID NO.:118, SEQ ID NO.:119, SEQ ID
NO.:120, SEQ ID NO.:121, SEQ ID NO.:122, SEQ ID NO.:123, SEQ ID NO.:124,
SEQ ID NO.:125, SEQ ID NO.:126, SEQ ID NO.:127. SEQ ID NO.:128, SEQ ID
NO.:129, SEQ ID NO.:130 and SEQ ID NO.:131.
[0481] Yet other aspects of the invention relate to a nucleic acid
encoding a heavy chain variable domain having at least 70%, 75%, 80%
sequence identity to a sequence selected from the group consisting of SEQ
ID NO.:18, SEQ ID NO.:22, SEQ ID NO.:26, SEQ ID NO.:132, SEQ ID NO.:133,
SEQ ID NO.:134, SEQ ID NO.:135, SEQ ID NO.:136, SEQ ID NO.:137, SEQ ID
NO.:138, SEQ ID NO.:139, SEQ ID NO.:140, SEQ ID NO.:141, SEQ ID NO.:142,
SEQ ID NO.:143, SEQ ID NO.:144, SEQ ID NO.:145, SEQ ID NO.:146, SEQ ID
NO.:147, SEQ ID NO.:148, SEQ ID NO.:149, SEQ ID NO.:150, SEQ ID NO.:151,
SEQ ID NO.:152, SEQ ID NO.:153, SEQ ID NO.:154, SEQ ID NO.:155, SEQ ID
NO.:156, SEQ ID NO.:157. Other aspects of the invention relates to the
use of a nucleic acid selected from the group consisting of SEQ ID NO.:1,
a fragment of 10 to 884 nucleotides of SEQ ID NO.:1 and a complement of
any of the preceding for impairing migration or survival of tumor cells
expressing KAAG1. Exemplary embodiments of such nucleic acid comprise
siRNAs, antisense, ribozymes and the like.
[0482] In yet another aspect, the present invention relates to a vector
comprising the nucleic acids described herein.
[0483] In accordance with the present invention, the vector may be an
expression vector.
[0484] Vector that contains the elements for transcriptional and
translational control of the inserted coding sequence in a particular
host are known in the art. These elements may include regulatory
sequences, such as enhancers, constitutive and inducible promoters, and
5' and 3' un-translated regions. Methods that are well known to those
skilled in the art may be used to construct such expression vectors.
These methods include in vitro recombinant DNA techniques, synthetic
techniques, and in vivo genetic recombination.
[0485] In another aspect the present invention relates to an isolated cell
which may comprise the nucleic acid described herein.
[0486] The isolated cell may comprise a nucleic acid encoding a light
chain variable domain and a nucleic acid encoding a heavy chain variable
domain either on separate vectors or on the same vector. The isolated
cell may also comprise a nucleic acid encoding a light chain and a
nucleic acid encoding a heavy chain either on separate vectors or on the
same vector.
[0487] In accordance with the present invention, the cell may be capable
of expressing, assembling and/or secreting an antibody or antigen binding
fragment thereof.
[0488] In another aspect, the present invention provides a cell which may
comprise and/or may express the antibody described herein.
[0489] In accordance with the invention, the cell may comprise a nucleic
acid encoding a light chain variable domain and a nucleic acid encoding a
heavy chain variable domain.
[0490] The cell may be capable of expressing, assembling and/or secreting
an antibody or antigen binding fragment thereof.
[0491] The examples below are presented to further outline details of the
present invention.
EXAMPLES
Example 1
[0492] This example describes the pattern of expression of the KAAG1 gene
in ovarian tumors and ovarian cancer cell line.
[0493] PCR analysis was performed to verify the percentage of ovarian
tumors that express the mRNA encoding KAAG1 (indicated as AB-0447 in the
Figure). The results showed that the KAAG1 gene is expressed in greater
than 85% of ovarian tumors from all stages of the disease and 100% of
late stage tumors. The expression of KAAG1 is lower or undetectable in
LMP samples (see FIG. 1A). For each sample, 1 .mu.g of amplified RNA was
reverse transcribed with random hexamers using an avian reverse
transcriptase, THERMOSCRIPT.TM. RT (Invitrogen). The cDNA was diluted and
1/200th of the reaction was used as template for each PCR reaction with
gene-specific primers as indicated. The primers used to amplify the KAAG1
mRNA contained the sequences shown in SEQ ID NOS:45 and 46. PCR reactions
were carried out in 96-well plates and half of the 25 .mu.l reaction was
electrophoresed on a 1% agarose gel. The gels were visualized and
photographed with a gel documentation system (BioRad). The upper panel of
FIG. 1A shows the results from 6 LMP samples (LMP) and 22 ovarian tumor
and 6 ovarian cell line (last 6 lanes on the right, OVCa) samples. The
lower panel of FIG. 1 shows the RNA samples from 30 normal tissues that
were tested as indicated.
[0494] KAAG1 expression was weakly detected in a few normal tissues
whereas the mRNA was evident in the fallopian tube and the pancreas (see
FIG. 1A). The amount of total RNA used in these reactions was controlled
with parallel PCR amplifications of glyceraldehyde-3-phosphate
dehydrogenase (GAPDH), a housekeeping gene, and the results showed that
equivalent starting material was present in each sample (see FIG. 1A).
The primers used to amplify the GAPDH gene contained the sequences shown
in SEQ ID NOs: 47 and 48. Thus, the expression of the KAAG1 gene fulfills
an important selection criteria: it is over-expressed in a large
proportion of ovarian tumors and its expression is low or absent in most
normal tissues. These data suggest that ovarian tumors may be
specifically targeted with high affinity monoclonal antibodies against
KAAG1.
[0495] Early stage cancer or tumors tend to be made up of cells that are
in a high state of differentiation but as the tumor progresses to a more
aggressive and invasive state, the cancer cells become increasingly
undifferentiated. There are needs to identify factors that contribute to
this transition and exploit these proteins as targets for the development
of therapeutics. Several ovarian cancer cell lines are available that
were derived from primary tumors and serve as excellent models for the
functional studies. The expression of KAAG1 was examined in these cell
lines. Four lines termed TOV-21G, TOV-112D, TOV-1946, and TOV-2223G were
established from primary tumors whereas OV-90 and OV-1946 are cell lines
derived from cells contained in ascites fluid of patients with advanced
ovarian cancer. Total RNA from cells established from primary tumors (see
in FIG. 1B, lanes 1, TOV-21G; 2, TOV-112D; 5, TOV-1946; 6, TOV-2223G) and
cells established from ascitic cells (lanes 3, OV-90; 4, OV-1946) was
converted to cDNA with reverse transcriptase and used as template in PCR
reactions with KAAG1-specific primers (SEQ ID NOS:45 and 46). As a
negative control, the reaction was carried out with total RNA from normal
ovary. Equal amounts of starting material were utilized as evidenced by
parallel PCR reactions with GAPDH (SEQ ID NOS:47 and 48). A sample of the
PCR reaction was electrophoresed on an agarose gel and visualized with
ethidium bromide. As shown in FIG. 1B, KAAG1 was detectable but weakly
expressed in the cell lines from the primary tumors and PCR reactions
performed at a higher number of cycles revealed the KAAG1 transcript in
all four of these cell lines. Conversely, both cell lines established
from the ascitic fluid cells exhibited high level of the KAAG1
transcript. The increased expression in cells from the ascitic fluid
suggests that the environment of the cells influences the regulation of
the KAAG1 gene.
[0496] Ascitic cells are associated with advanced disease and the pattern
of expression disclosed in FIG. 1B implies that increased KAAG1 levels
are associated with anchorage-independent growth. This question was
addressed by culturing the cells in hanging droplets, a condition that
prevents the cells from adhering to the petri dish, as is the case when
they are grown as monolayers. These so called three-dimensional cultures
allow the cells to associate and the formation of spheroids is observed
(see FIG. 1C). Spheroids were cultures as follows: TOV-112D, OV-90, or
TOV-21G cells (4 000 in 15 .mu.l) were incubated for 4 days in medium in
the absence (left panels, FIG. 1C) or presence of 5% FBS (right panels,
FIG. 1C, +5% serum). The magnification of the image was set to
100.times.. These spheroids have been extensively characterized and
exhibit many of the properties found in primary tumors including
morphological and functional properties as well as the molecular
signature as measured by microarray-based expression profiling.
[0497] Total RNA was isolated from spheroid preparations and RT-PCR was
performed as described for FIG. 1A. TOV-21G, TOV-112D, OV-90 cells were
seeded as described in the legend for FIG. 1C under conditions to produce
spheroids. After 4 days, total RNA was isolated and used to perform
RT-PCR reactions with KAAG1-specific primers (SEQ ID NOS:45 and 46). PCR
reactions were electrophoresed on agarose gels. Conducting parallel
reactions to amplify GAPDH (SEQ ID NOS:47 and 48) demonstrated that equal
amounts of starting material were present in each sample. The following
acronyms are used in FIG. 1D: Ce., cells grown as monolayers; Sph., cells
grown as spheroids. Strikingly, KAAG1 expression was up-regulated when
TOV-21G and TOV-112D were grown as spheroids (see FIG. 1D). In the case
of the OV-90 cells, the level of expression of the KAAG1 gene was
unchanged and remained very high. Presumably, the level of expression
attained in the cell lines derived from the ascitic fluid, as exemplified
by the OV-90 cells and the OV-1946 cells (see FIG. 1A) has reached a
maximum.
[0498] These results correlated with the previous data showing high
expression in cell lines derived from ascitic fluid and confirm that
expression of KAAG1 is influenced by the microenvironment of the cancer
cells. Additionally, the up-regulation of KAAG1 transcription that was
observed in spheroids implies that high levels of KAAG1 are present in
malignant ovarian cancer.
Example 2
[0499] This example describes in vitro results that suggest a critical
role for KAAG1 in the survival of ovarian cancer cells.
[0500] With the demonstration that KAAG1 expression is regulated in
ovarian cancer cells, the function of this gene in these cells was
examined. To address this question, in vitro assays were conducted to
determine if this protein plays a role in cancer cell proliferation,
migration, and/or survival. RNAi was used to knock down the expression of
the endogenous KAAG1 gene in the TOV-21G ovarian cancer cell line. The
design of two separate short-hairpin RNA (shRNA) sequences was performed
using web-based software that is freely available to those skilled in the
art (Qiagen for example). These chosen sequences, usually 19-mers, were
included in two complementary oligonucleotides that form the template for
the shRNAs, i.e. the 19-nt sense sequence, a 9-nt linker region (loop),
the 19-nt antisense sequence followed by a 5-6 poly-T tract for
termination of the RNA polymerase III. The sequences of the 19-mers that
were used to knock down the expression of KAAG1 are shown in SEQ ID
NOS:49 and 50. Appropriate restriction sites were inserted at the ends of
these oligonucleotides to facilitate proper positioning of the inserts so
that the transcriptional start point is at a precise location downstream
of the hU6 promoter. The plasmid utilized in all RNA interference
studies, pSilencer 2.0 (SEQ ID NO.:51), was purchase from a commercial
supplier (Ambion, Austin, Tex.). Two different shRNA expression vectors
were constructed to increase the chance of observing RNAi effects and the
specificity of phenotypic observations. TOV-21G cells were seeded in
6-well plates and transfected 24 h later with 1 .mu.g of pSil-shRNA
vector. Sh.1 and sh.2 were used to designate 2 different shRNA sequences
targeting the KAAG1 gene. Stable transfectants were selected for 5-7
days, expanded, and grown to confluence. All of the following in vitro
cell-based assays were performed using these stably transfected cell
lines that contain shRNAs specific for KAAG1.
[0501] The migration or mobility of the cells was measured in a standard
cell motility assay. This scratch assay, as it is called, measures the
speed at which cells fill a denuded area in a confluent monolayer. As
illustrated in FIG. 2A, TOV-21G cells containing the scrambled shRNA
filled up the wound almost completely after 24 h compared to the control
untreated cells (compare middle-left panel with left panel). By contrast,
the ability of TOV-21G cells expressing KAAG1 shRNAs to fill the denuded
area was greatly reduced. In fact, the number of cells that filled the
denuded area in the presence of the KAAG1 shRNA cells more closely
resembled the number of cells at time 0 h (compare the left panel with
the right panels).
[0502] To examine the longer-term effects of reduced expression of KAAG1
in ovarian cancer cells, the cells were extensively diluted and cultured
for 10 days in a colony survival assay. TOV-21G cells were seeded in
12-well plates at a density of 50 000 cells/well and transfected 24 h
later with 1 .mu.g of pSil-shRNA vector. Sh-1 and sh-2 are used to
designate 2 different shRNA sequences targeting the same gene. The next
day, fresh medium was applied containing 2 .mu.g/ml puromycin and the
selection of the cells was carried out for 3 days. The cells were washed
and fresh medium without puromycin was added and growth continued for
another 5 days. To visualize the remaining colonies, the cells were
washed in PBS and fixed and stained simultaneously in 1% crystal
violet/10% ethanol in PBS for 15 minutes at room temperature. Following
extensive washing in PBS, the dried plates were scanned for photographic
analysis. A significant decrease in the survival of the cancer cell line
was observed and a representative experiment is displayed in FIG. 2B.
Identical results were obtained when the shRNAs were transfected into
another ovarian cancer cell line, TOV-112D.
[0503] Thus, taken together, the regulated expression of KAAG1 in detached
cells coupled with the requirement of this gene in the migration and the
survival of ovarian cancer cells supports an important role for KAAG1 in
ovarian cancer cells. Furthermore, these experiments suggest that an
antagonist of KAAG1 protein, such as a monoclonal antibody, would result
in reduced invasiveness and decreased tumor survival.
Example 3
[0504] This example provides details pertaining to the family of
monoclonal antibodies that bind to KAAG1.
[0505] The antibodies that bind KAAG1 were generated using the Biosite
phage display technology. A detailed description of the technology and
the methods for generating these antibodies can be found in the U.S. Pat.
No. 6,057,098. Briefly, the technology utilizes stringent panning of
phage libraries that display the antigen binding fragments (Fabs). After
a several rounds of panning, a library, termed the OMNICLONAL.TM., was
obtained that was enriched for recombinant Fabs containing light and
heavy chain variable regions that bound to KAAG1 with very high affinity
and specificity. From this library, more precisely designated
OMNICLONAL.TM. AL0003Z1, 96 individual recombinant monoclonal Fabs were
prepared from E. coli and tested for KAAG1 binding.
[0506] To measure the relative binding of each individual monoclonal
antibody, recombinant human KAAG1 was produced in 293E cells using the
large-scale transient transfection technology (Durocher et al., 2002;
Durocher, 2004). The entire coding region of the KAAG1 cDNA was amplified
by PCR using a forward primer that incorporated a BamHI restriction site
(SEQ ID NO.:52) and a reverse primer that incorporated a HindIII
restriction site (SEQ ID NO.:53). The resulting PCR product measured 276
base pairs and following digestion with BamHI and HindIII, the fragment
was ligated into the expression vector pYD5 (SEQ ID NO.:54) that was
similarly digested with the same restriction enzymes. The pYD5 expression
plasmid contains the coding sequence for the human Fc domain that allows
fusion proteins to be generated as well as the sequence encoding the IgG1
signal peptide to allow the secretion of the fusion protein into the
culture medium. For each milliliter of cells, one microgram of the
expression vector, called pYD5-0447, was transfected in 293E cells grown
in suspension to a density of 1.5-2.0 million cells/ml. The transfection
reagent used was polyethylenimine (PEI), (linear, MW 25,000, Cat#23966
Polysciences, Inc., Warrington, Pa.) which was included at a DNA:PEI
ratio of 1:3. Growth of the cells was continued for 5 days after which
the culture medium was harvested for purification of the recombinant
Fc-KAAG1 fusion protein. The protein was purified using Protein-A agarose
as instructed by the manufacturer (Sigma-Aldrich Canada Ltd., Oakville,
ON). A representative polyacrylamide gel showing a sample of the purified
Fc-KAAG1 (indicated as Fc-0447) is shown in FIG. 3A.
[0507] The 96-well master plate of monoclonal preparations contained
different concentrations of purified anti-KAAG1 Fabs in each well. A
second stock master plate was prepared by diluting the Fabs to a final
concentration of 10 .mu.g/ml from which all subsequent dilutions were
performed for ELISA measurements. To carry out the binding of Fc-KAAG1 to
the monoclonal preparations, the Fc-KAAG1 was biotinylated with
NHS-biotin (Pierce, Rockford, Ill.) and 10 ng/well was coated in a
streptavidin 96-well plate. One nanogram of each Fab monoclonal
preparation was added to each well and incubated at room temperature for
30 minutes. Bound antibody was detected with HRP-conjugated mouse
anti-kappa light chain antibody in the presence of TMB liquid substrate
(Sigma-Aldrich Canada Ltd., Oakville, ON) and readings were conducted at
450 nm in microtiter plate reader. As shown in FIG. 3B, a total of 48
(highlighted in grey) monoclonal antibodies displayed significant binding
in this assay (>0.1 arbitrary OD.sub.450 units). The antibodies were
purposely diluted to 1 ng/well to accentuate the binding of those
antibodies with the most affinity for KAAG1. As a control, the antibodies
did not bind to biotinylated Fc domain. These data also revealed that the
binding of the antibodies varied from well to well indicating that they
exhibited different affinities for KAAG1.
Example 4
[0508] This example describes the epitope mapping studies to determine
which region of KAAG1 the antibodies bind to.
[0509] To further delineate the regions of KAAG1 that are bound by the
monoclonal antibodies, truncated mutants of KAAG1 were expressed and used
in the ELISA. As for the full length KAAG1, the truncated versions were
amplified by PCR and ligated into BamHI/HindIII digested pYD5. The
primers that were used combined the forward oligonucleotide with the
sequence shown in SEQ ID NO.:52 with primers of SEQ ID NOS:55 and 56, to
produce Fc-fused fragments that ended at amino acid number 60 and 35 of
KAAG1, respectively. The expression of these mutants was conducted as was
described above for the full length Fc-KAAG1 and purified with Protein-A
agarose. A representative gel of the protein preparations that were used
in the ELISA is shown in FIG. 4A and a schematic of the mutant proteins
used for epitope mapping is depicted in FIG. 4B.
[0510] The results showed that the library was comprised of antibodies
that could bind to each of the delineated KAAG1 regions. In particular,
of the 48 mAbs that bound to KAAG1 in the first ELISA, nine (wells A2,
A12, C2, C4, D1, E10, F1, H3, and H8) were found to interact with the
first 35 amino acids of KAAG1 whereas five (D12, E8, F5, G10, and H5)
were found to interact with the last 25 amino acids of KAAG1. Thus, the
remaining 34 antibodies interacted with a region of KAAG1 spanned by
amino acids 36-59. These results were in agreement with the sequence
analysis of 24 representative light and heavy chain variable regions.
Indeed, alignment of these sequences revealed that the antibodies
clustered into three groups based on the percentage identity in their
respective CDRs. Antibodies contained in each cluster all interacted with
the same region of KAAG1.
[0511] Therefore, based on the relative binding affinity of the mAb,
differential epitope interaction characteristics, and the differences in
variable domain sequences, three antibodies from the plate described in
Example 3 were selected for further analysis as exemplary anti-KAAG1
monoclonal antibodies.
Example 5
[0512] This example discloses the methods used to convert the Fabs into
full IgG1 chimeric monoclonal antibodies. A scheme of the methodology is
presented in FIG. 5.
[0513] Aside from the possibility of conducting interaction studies
between the Fab monoclonals and the KAAG1 protein, the use of Fabs is
limited with respect to conducting meaningful in vitro and in vivo
studies to validate the biological function of the antigen. Thus, it was
necessary to transfer the light and heavy chain variable regions
contained in the Fabs to full antibody scaffolds, to generate mouse-human
chimeric IgG1s. The expression vectors for both the light and heavy
immunoglobulin chains were constructed such that i) the original
bacterial signal peptide sequences upstream of the Fab expression vectors
were replaced by mammalian signal peptides and ii) the light and heavy
chain constant regions in the mouse antibodies were replaced with human
constant regions. The methods to accomplish this transfer utilized
standard molecular biology techniques that are familiar to those skilled
in the art. A brief overview of the methodology is described here (see
FIG. 5).
[0514] Light Chain Expression Vector--
[0515] An existing mammalian expression plasmid, called pTTVH8G (Durocher
et al., 2002), designed to be used in the 293E transient transfection
system was modified to accommodate the mouse light chain variable region.
The resulting mouse-human chimeric light chain contained a mouse variable
region followed by the human kappa constant domain. The cDNA sequence
encoding the human kappa constant domain was amplified by PCR with
primers OGS1773 and OGS1774 (SEQ ID NOS:57 and 58, respectively). The
nucleotide sequence and the corresponding amino acid sequence for the
human kappa constant region are shown in SEQ ID NOS:59 and 60,
respectively. The resulting 321 base pair PCR product was ligated into
pTTVH8G immediately downstream of the signal peptide sequence of human
VEGF A (NM_003376). This cloning step also positioned unique restriction
endonuclease sites that permitted the precise positioning of the cDNAs
encoding the mouse light chain variable regions. The sequence of the
final expression plasmid, called pTTVK1, is shown in SEQ ID NO.:61. Based
on the sequences disclosed in Table 3, PCR primers specific for the light
chain variable regions of antibodies 3D3, 3G10, and 3C4 (SEQ ID NOS:15,
19, and 23, respectively) were designed that incorporated, at their
5'-end, a sequence identical to the last 20 base pairs of the VEGF A
signal peptide. The sequences of these primers are shown in SEQ ID
NOS:62, 63, and 64. The same reverse primer was used to amplify all three
light chain variable regions since the extreme 3'-ends were identical.
This primer (SEQ ID NO.:65) incorporated, at its 3'-end, a sequence
identical to the first 20 base pairs of the human kappa constant domain.
Both the PCR fragments and the digested pTTVK1 were treated with the
3'-5' exonuclease activity of T4 DNA polymerase resulting in
complimentary ends that were joined by annealing. The annealing reactions
were transformed into competent E. coli and the expression plasmids were
verified by sequencing to ensure that the mouse light chain variable
regions were properly inserted into the pTTVK1 expression vector. Those
skilled in the art will readily recognize that the method used for
construction of the light chain expression plasmids applies to all
anti-KAAG1 antibodies contained in the original Fab library.
[0516] Heavy Chain Expression Vector--
[0517] The expression vector that produced the heavy chain immunoglobulins
was designed in a similar manner to the pTTVK1 described above for
production of the light chain immunoglobulins. Plasmid pYD11 (Durocher et
al., 2002), which contains the human IgGK signal peptide sequence as well
as the CH2 and CH3 regions of the human Fc domain of IgG1, was modified
by ligating the cDNA sequence encoding the human constant CH1 region. PCR
primers OGS1769 and OGS1770 (SEQ ID NOS:66 and 67), designed to contain
unique restriction endonuclease sites, were used to amplify the human
IgG1 CH1 region containing the nucleotide sequence and corresponding
amino acid sequence shown in SEQ ID NOS:68 and 69. Following ligation of
the 309 base pair fragment of human CH1 immediately downstream of the
IgGK signal peptide sequence, the modified plasmid (SEQ ID NO.:70) was
designated pYD15. When a selected heavy chain variable region is ligated
into this vector, the resulting plasmid encodes a full IgG1 heavy chain
immunoglobulin with human constant regions. Based on the sequences
disclosed in Table 3, PCR primers specific for the heavy chain variable
regions of antibodies 3D3, 3G10, and 3C4 (SEQ ID NOS:17, 21, and 25,
respectively) were designed that incorporated, at their 5'-end, a
sequence identical to the last 20 base pairs of the IgGK signal peptide.
The sequences of these primers are shown in SEQ ID NOS:71 (3D3 and 3G10
have the same 5'-end sequence) and 72. The same reverse primer was used
to amplify all three heavy chain variable regions since the extreme
3'-ends were identical. This primer (SEQ ID NO.:73) incorporated, at its
3'-end, a sequence identical to the first 20 base pairs of the human CH1
constant domain. Both the PCR fragments and the digested pYD15 were
treated with the 3'-5' exonuclease activity of T4 DNA polymerase
resulting in complimentary ends that were joined by annealing. The
annealing reactions were transformed into competent E. coli and the
expression plasmids were verified by sequencing to ensure that the mouse
heavy chain variable regions were properly inserted into the pYD15
expression vector. Those skilled in the art will readily recognize that
the method used for construction of the heavy chain expression plasmids
applies to all anti-KAAG1 antibodies contained in the original Fab
library.
[0518] Expression of Human IgG1s in 293E Cells--
[0519] The expression vectors prepared above that encoded the light and
heavy chain immunoglobulins were expressed in 293E cells using the
transient transfection system (Durocher et al., 2002). The methods used
for co-transfecting the light and heavy chain expression vectors were
described in Example 3. The ratio of light to heavy chain was optimized
in order to achieve the most yield of antibody in the tissue culture
medium and it was found to be 9:1 (L:H). The ability of the chimeric
anti-KAAG1 monoclonal antibodies to bind to recombinant Fc-KAAG1 was
measured in the ELISA and compared with the original mouse Fabs. The
method was described in Example 3. As depicted in FIG. 6, the binding of
the 3D3, and 3G10 chimeric IgG1 monoclonal antibodies was very similar to
the Fabs. In the case of the 3C4, the binding activity of the chimeric
was slightly less than the Fab. Despite this, this result shows that the
transposition of the variable domains from the mouse Fabs into a human
IgG1 backbone did not significantly affect the capacity of the light and
heavy chain variable regions to confer KAAG1 binding.
Example 6
[0520] This example describes the use of anti-KAAG1 antibodies to block
the activity of KAAG1 in ovarian cancer cell models.
[0521] Example 2 disclosed RNAi studies showing that KAAG1 played an
important role in the behavior of ovarian cancer cells. The monoclonal
antibodies described above were used to determine whether it was possible
to reproduce these results by targeting KAAG1 at the cell surface.
TOV-21G and OV-90 cells were cultured under conditions to produce
spheroids and treated with 10 .mu.g/ml of 3D3, 3G10, or 3C4 anti-KAAG1
chimeric monoclonal antibody. As illustrated in FIG. 7, both cell lines
efficiently formed spheroids when left untreated (parental) or when
treated with antibody dilution buffer (control). In contrast, the
presence of anti-KAAG1 antibodies resulted in loosely packed structures
and in certain cases, the cells were unable to assemble into spheroids.
These results confirm the earlier observations and suggest that the
anti-KAAG1 monoclonal antibodies can modulate the activity of KAAG1
during the formation of spheroids. Since spheroid formation by cancer
cell lines is an in vitro model for tumor formation, the results also
suggest that blocking KAAG1 could lead to decreased tumor formation in
vivo.
Example 7
[0522] This example describes the use of anti-KAAG1 antibodies for
detecting the expression of KAAG1 in ovarian tumors.
[0523] As a means of confirming the expression of KAAG1 protein in ovarian
cancer tumors and in order determine if expression of the gene correlated
with the presence of the protein, immunohistochemistry was conducted.
Tissue microarrays were obtained that contained dozens of ovarian tumor
samples generated from patient biopsies. Paraffin-embedded epithelial
ovarian tumor samples were placed on glass slides and fixed for 15 min at
50.degree. C. Deparaffinization was conducted by treating 2.times. with
xylene followed by dehydration in successive 5 min washes in 100%, 80%,
and 70% ethanol. The slides were washed 2.times. in PBS for 5 min and
treated with antigen retrieval solution (citrate-EDTA) to unmask the
antigen. Endogenous peroxide reactive species were removed by incubating
slides with H.sub.2O.sub.2 in methanol and blocking was performed by
incubating the slides with serum-free blocking solution (Dakocytomation)
for 20 min at room temperature. The primary mAb (anti-KAAG1 3D3) was
added for 1 h at room temperature. KAAG1-reactive antigen was detected by
incubating with biotin-conjugated mouse anti-kappa followed by
streptavidin-HRP tertiary antibody. Positive staining was revealed by
treating the slides with DAB-hydrogen peroxide substrate for less than 5
min and subsequently counterstained with hematoxylin. The KAAG1 protein
was found to be expressed at very high levels in the vast majority of
ovarian tumor samples. A representative array containing 70 tumors is
depicted in FIG. 8A. As demonstrated by the expression profiling studies
that were performed using RT-PCR, KAAG1 transcripts were present in
greater than 85% of ovarian tumor samples analyzed. Clearly, there is an
excellent correlation between the transcription of the KAAG1 gene and the
presence of the protein in ovarian cancer. Some of the samples were
inspected at a higher magnification to determine which cells were
expressing the KAAG1 protein. As depicted in FIG. 8B, KAAG1 is
predominantly expressed in the surface epithelium of ovarian tumors. In
addition, strong intensity was observed on the apical side of these
epithelial cells (see arrows in FIG. 8B, magnification: 20.times.).
Finally, immunohistochemistry was repeated on ovarian tumor samples that
originated from different histotypes. As explained earlier, epithelial
ovarian cancer can be classified into 4 major histotypes: serous,
endometroid, clear cell, and mucinous. The expression of KAAG1 was
detected in all types of epithelial ovarian cancer, in particular serous
and endometroid histotypes (see FIG. 8C).
[0524] Taken together, these immunohistochemical studies illustrate the
utility of detecting KAAG1 in ovarian cancer with the monoclonal
antibodies.
Example 8
IgG.sub.1 Antibodies Against KAAG1 can Mediate ADCC
[0525] Antibody-Dependent Cell Cytotoxicity (ADCC) is a mechanism of
cell-mediated immunity whereby effector cells, typically natural killer
(NK) cells, of the immune system actively lyse target cells that have
been bound by specific antibodies. The interaction between the NK cells
and the antibody occurs via the constant Fc domain of the antibody and
high-affinity Fc.gamma. receptors on the surface of the NK cells.
IgG.sub.1s have the highest affinity for the Fc receptors while IgG.sub.2
mAbs exhibit very poor affinity. For this reason the chimeric antibodies
targeting KAAG1 were designed as IgG.sub.1s. This type of effector
function that is mediated in this manner can often lead to the selective
killing of cancer cells that express high level of antigen on their cell
surfaces.
[0526] An in vitro assay to measure ADCC activity of the anti-KAAG1
IgG.sub.1 chimeric antibodies was adapted from a previously published
method, which measured the ADCC activity of the anti-CD20 rituxan in the
presence of a lymphoma cell line called WIL2-S (Idusogie et al., (2000)
J. Immunol. 164, 4178-4184). Human peripheral blood mononuclear cells
(PBMNCs) were used as a source of NK cells which were activated in the
presence of increasing concentration of the 3D3 chimeric IgG.sub.1
antibody (FIG. 13). The target cells were incubated with the activated
PBMNCs at a ratio of 1 to 25. As shown, cell death increased in a
dose-dependent manner both in the presence of OVCAR-3 and the lymphoma
cell line, the latter of which was shown to express KAAG1 by RT-PCR (not
shown). As a positive control, the results from the published method were
reproduced where high level of ADCC was obtained for rituxan in the
presence of WIL2-S cells.
[0527] ADCC was also observed with other ovarian cancer cell lines that
express relatively high levels of KAAG1. These results demonstrate that
IgG.sub.1 antibodies that are specific for KAAG1, as exemplified by 3D3,
can enhance the lysis of cancer cells which express the antigen on their
cell surface.
Example 9
Antibodies Against KAAG1 can Reduce the Invasion of Ovarian Tumors
[0528] Patients that develop ovarian cancer have lesions that typically
initiate by an uncontrolled growth of the cells in the epithelial layer
of the ovary or, in some instances, the fallopian tube. If detected
early, these primary tumors are surgically removed and first-line
chemotherapy can result in very good response rates and improved overall
survival. Unfortunately, 70% of the patients will suffer recurrent
disease resulting in the spread of hundreds of micro-metastatic tumors
throughout the abdominal cavity. Second-line therapies can be
efficacious, but often patients either respond poorly or the tumors
develop chemoresistance. Treatment options are limited and there are
urgent needs for new therapies to circumvent resistance to cytotoxic
drugs.
[0529] In order to test the efficacy of anti-KAAG1 antibodies in vivo, an
animal model of ovarian cancer was used that is the closest
representation of the clinical manifestation of the disease in humans.
The TOV-112D cell line is of endometrioid origin and expresses the KAAG1
antigen as measured by RT-PCR. Previous IHC studies showed that ovarian
tumors of the endometrioid histotype contain strong expression of KAAG1
thus rendering the 112D cell line an appropriate selection for testing
anti-KAAG1 antibodies.
[0530] 1.times.10.sup.6 cells were implanted in the peritoneal cavity of
SCID mice in a volume of 200 .quadrature.L. Treatment with either PBS or
antibodies diluted in PBS was performed 2 days later at a dose of 25
mg/kg qwk. The mice were sacrificed as soon as the tumors were detected
by palpation of the abdomen. The intra-peritoneal inoculation of the
TOV-112D cell line in SCID mice resulted in the implantation of dozens of
micro-metastatic tumors that closely resemble those that are observed in
humans. Mice treated with PBS, the diluent for the antibodies, contained
upon examination, an average of 25-30 tumors per animal (FIGS. 14A and
14B). In some cases, the number of tumors was so high in the abdominal
cavity of these mice that the number of tumors could not be easily
determine; these mice were excluded from the statistical analysis. When
the mice were treated with the 3C4 and 3D3 antibodies, the number of
micro-metastatic tumors was drastically reduced. In addition, there was
at least one animal per group treated with anti-KAAG1 where no tumors
were seen. A second experiment was conducted in mice containing a larger
number of TOV-112D tumors (>50/animal) and very similar results were
obtained. Moreover, there was very little difference between the groups
treated with the 3C4 compared to the 3D3 antibody. However, the tendency
in these in vivo experiments as well as the results obtained in the
cell-based assays show that the 3D3 antibody displayed slightly more
efficacy. Whether, this is due to a more accessible epitope or a higher
affinity of 3D3 compared to 3C4 for the antigen still remains to be
established. The results from these two experiments demonstrated that
targeting KAAG1 on the surface of ovarian cancer cells could lead to a
significant reduction in the spread of the tumors in vivo. The anti-KAAG1
mAbs prevent the spread of TOV-112D ovarian tumors in vivo.
[0531] Furthermore, these findings are in complete agreement with the
observations that were made in the cell-based assays. For example, the
increased expression of the KAAG1 mRNA in the spheroids compared to cell
lines grown as monolayers; the reduction in cell migration in the
presence of KAAG1 shRNAs, the reduction in the ability of cell lines to
form spheroids when treated with KAAG1 antibodies; and finally,
enhancement of ADCC activity by anti-KAAG1 IgG.sub.1s. Taken together,
the results strongly suggest that targeting KAAG1 with an antibody has
great therapeutic potential in recurrent ovarian cancer.
Example 10
[0532] KAAG1 is Expressed in Skin Tumors and Renal Cell Carcinomas and is
a Therapeutic Target in these Indications.
[0533] The mRNA profiling studies that were conducted showed that the
transcript encoding the KAAG1 antigen was highly expressed in cell lines
derived from melanoma samples and renal carcinomas. These results were
disclosed in Sooknanan et al., 2007. To confirm the transcriptional
regulation of the KAAG1 gene in these cancer types, immunohistochemistry
was performed with an anti-KAAG1 antibody on human skin tumor tissue
microarrays (Pantomics Inc., Richmond, Calif.) containing several
sections isolated from squamous cell carcinomas and melanomas. The
analysis of this array showed that there was very strong staining in
biopsies isolated from squamous cell carcinomas and melanomas (FIG. 15,
top panel). Both of these types are among the most common forms of skin
cancers and interestingly, the squamous cell carcinomas are the most
metastatic, a fact that again links the expression of KAAG1 to an
invasive phenotype. As previously observed, the presence of KAAG1 was
very weak or absent on the three normal skin samples that were contained
on the array. Similarly, KAAG1 was detected in many of the samples
contained in an array of renal cancer. Most of the positive samples were
predominantly of the papillary cell carcinoma type and a few clear cell
carcinomas expressed KAAG1 protein. Papillary carcinomas represent
approximately 20% of renal cancer cases.
[0534] In order to test if the function of KAAG1 is the same in these
types of cancer compared to its role in ovarian cancer, cell lines
derived from melanoma and renal cell carcinomas were obtained and tested
in the spheroid culture assay (see Example 1 and 6). For the melanoma
model, A375 and SK-MEL5 cells, two malignant melanoma cell lines, were
cultured under conditions that allowed them to form spheroids in the
presence of 5% FBS. The cultures were incubated with or without the
anti-KAAG1 chimeric 3D3 antibody at a concentration of 5 .mu.g/ml. As
shown in FIG. 16, inclusion of 3D3 antibody in the cultures prevented the
proper assembly of spheroid structures in melanoma cell lines. This
result suggested that KAAG1 plays a similar role in melanoma as it does
in ovarian cancer. Cell lines derived from renal cell carcinoma were also
tested. The A-498 cell line is a renal papillary cell carcinoma cell line
whereas the 786-O is a renal clear cell carcinoma. As depicted in FIG.
16, only the A-498 spheroids were affected by the presence of the 3D3
anti-KAAG1 antibody while the 786-O cell line was unaffected in this
assay. These results parallel the immunohistochemistry results described
above and indicate that the inhibition of spheroids formation is
dependent on the presence of KAAG1 on the surface of renal cancer cells
derived predominantly from papillary kidney cancers. It is possible
however, that the anti-KAAG1 antibody may work in other types of assays
for renal clear cell carcinoma.
[0535] Taken together, these data are strongly supportive of a critical
function in role of KAAG1 in melanoma and kidney cancer and indicate that
blocking KAAG1 with antibodies in these indications has therapeutic
potential.
Example 11
KAAG1 is Expressed on the Surface of Ovarian Cancer Cells.
[0536] The combined results from the bioinformatics analysis of the
primary structure of the cDNA encoding KAAG1, biochemical studies, and
immunohistochemical detection of the protein in epithelial cells
suggested that the KAAG1 antigen was located on the cell surface.
However, more direct evidence was required to demonstrate that KAAG1 is
indeed a membrane-bound protein. In one approach, ovarian cancer cell
lines known to express KAAG1 were plated in micro-titer plates, fixed
under conditions that do not permeate the cells, and incubated with
increasing concentration of anti-KAAG1 chimeric antibodies. Following
extensive washing of the cells, bound antibody was detected with
HRP-conjugated anti-human IgG as a secondary antibody in a modified
cell-based ELISA (see FIG. 17A). The first observation that can be made
from these experiments is that the antibodies could be specifically
captured by the cells suggesting that the KAAG1 was present at the cell
surface. Secondly, the amount of binding was strongest on SKOV-3 cells
and the TOV-21G cells exhibited the weakest binding. This was in complete
agreement with RT-PCR data which demonstrated that the KAAG1 mRNA was
expressed in similar proportions in these cell lines (not shown).
Additionally, the 3D3 antibody produced the strongest signal implying
that the epitope targeted by this antibody was the most accessible in
this assay. The 3G10 could only detect KAAG1 in the cell line that
expressed the highest level of AB-0447 (SKOV-3 cells, see right panel of
FIG. 17A). A second approach used was flow cytometry. In this case, a
mouse 3D3 anti-KAAG1 antibody was incubated with SKOV-3 ovarian cancer
cells at saturating conditions and following extensive washing, the bound
3D3 anti-KAAG1 antibody was detected with anti-mouse IgG conjugated to
FITC in a flow cytometer. As shown in FIG. 17B, the signal at the surface
of SKOV-3 cells was much higher compared to same cells labeled with the
negative control, an anti-KLH (Keyhole limpet hemocyanin) antibody,
specific for a non-mammalian unrelated protein, which was at a
fluorescence level the same as the background readings. Taken together,
these results demonstrate that KAAG1 is located on the surface of cells.
Example 12
Methods for the Use of Humanized Anti-KAAG1 Antibodies.
[0537] On the basis of both the in vitro and preliminary in vivo results,
two mouse anti-KAAG1 antibody candidates, designated 3D3 and 3C4, were
selected for humanization using in silico modeling using methods familiar
to those in the art. In brief, the variable regions of the murine
antibodies were modeled in 3D based on available crystal structures of
mouse, humanized, and fully human variable regions that displayed high
sequence homology and similar CDR loop lengths. The CDRs are the amino
acid sequences that contribute to antigen binding; there are 3 CDRs on
each antibody chain. Additionally, the framework regions, the amino acid
sequences that intervene between the CDRs, were modified by standard
homology comparison between mouse and human antibody sequences resulting
in the `best-fit` human sequence. These modifications ensured that the
proper positioning of the CDR loops was maintained to ensure maximum
antigen binding in the humanized structure as well as preserving the
potential N- and O-linked glycosylation sites. The sequence of both the
heavy and light chain variable regions in the humanized (h) 3D3 and 3G4
resulted in 96% and 94% humanization, respectively. The structure of the
3D3 and 3C4 models for each antibody is shown in FIGS. 18A and 18B,
respectively. As illustrated in these structures, the 3D3 required the
maintenance of 3 unusual amino acids (FIG. 18A, Met93 and Gly94 on the
heavy chain and Ser57 on the light chain) because of their proximity to
the CDRs. Modeling predicted that replacement of these mouse amino acids
with human equivalents might compromise binding of the antibody with the
KAAG1 antigen. In the case of 3C4, 6 amino acids were considered unusual
(FIG. 18B, Glu1, Gln72 and Ser98 on the heavy chain and Thr46, Phe49 and
Ser87 on the light chain). In both figures, the light chain CDRs are
indicated by L1, L2, and L3 for CDR1, CDR2, and CDR3, respectively,
whereas the heavy chain CDRs are indicated by H1, H2, and H3 for CDR1,
CDR2, and CDR3, respectively.
[0538] The sequences that encode the complete anti-KAAG1 3D3
immunoglobulin light and heavy chains are shown in SEQ ID NO.:176 and
177, respectively. The variable region of the humanized 3D3 light chain
is contained between amino acids 21-133 of SEQ ID NO.:176 and is shown in
SEQ ID NO.:178. The variable region of the humanized 3D3 heavy chain is
contained between amino acids 20-132 of SEQ ID NO.:177 and is shown in
SEQ ID NO.:179. The sequences that encode the complete anti-KAAG1 3C4
immunoglobulin light and heavy chains are shown in SEQ ID NO.:180 and
181, respectively. The variable region of the humanized 3C4 light chain
is contained between amino acids 21-127 of SEQ ID NO.:180 and is shown in
SEQ ID NO.:182. The variable region of the humanized 3C4 heavy chain is
contained between amino acids 19-136 of SEQ ID NO.:181 and is shown in
SEQ ID NO.:183.
[0539] Following assembly of expression vectors and production of the h3D3
in transfected mammalian cells (see Example 5), several assays were
performed to demonstrate the bio-equivalence of the humanization process.
Since an antibody harboring effector functions was required, the h3D3 was
assembled as a human IgG.sub.1. ELISA-based assays were performed to
directly compare the ability of the h3D3 to recombinant KAAG1. The
methods used to perform these tests were as described in Example 3 using
recombinant Fc-KAAG1. As shown in FIG. 19A, the binding activity of the
h3D3 was identical to that of the chimeric 3D3.
[0540] More precise measurements were conducted using Surface Plasmon
Resonance (SPR) in a BIACORE.TM. instrument. Kinetic analysis was used to
compare the affinity of the chimeric 3D3 with the h3D3 as well as with
hybrid antibodies encompassing different permutations of the light and
heavy chains (see FIG. 19B). Briefly, anti-human Fc was immobilized on
the BIACORE.TM. sensor chip and chimeric or h3D3 was captured on the
chip. Different concentrations of monomeric recombinant KAAG1 were
injected and the data were globally fitted to a simple 1:1 model to
determine the kinetic parameters of the interaction. The kinetic
parameters of the chimeric 3D3 were tabulated in FIG. 19B (m3D3). The
average K.sub.D of the chimeric 3D3 was 2.35.times.10.sup.-10 M. In
comparison, all permutations of the chimeric (C)/humanized (H) displayed
very similar kinetic parameters. The average K.sub.D of the chimeric
light chain expressed with the chimeric heavy chain (indicated as `CC` in
FIG. 19B) was 2.71.times.10.sup.-10 M, the average K.sub.D of the
humanized light chain expressed with the chimeric heavy chain (indicated
as `HC` in FIG. 19B) was 3.09.times.10.sup.-10 M, the average K.sub.D of
the chimeric light chain expressed with the humanized heavy chain
(indicated as `CH` in FIG. 19B) was 5.05.times.10.sup.-10 M, and the
average K.sub.D of the humanized light chain expressed with the humanized
heavy chain (indicated as `HH` in FIG. 19B) was 4.39.times.10.sup.-10 M.
The analyses indicated that the humanization of 3D3 conserved the binding
activity of the original mouse antibody.
[0541] The biological function of the h3D3 was evaluated in the spheroid
culture assay (see Example 6). SKOV-3 ovarian cancer cells were cultured
in the presence of 5% FBS in the presence of h3D3 or a non-KAAG1 binding
isotype control antibody. The results (shown in FIG. 19C), indicated that
treatment with either the buffer or the non-related IgG did not inhibit
the formation of the compact 3-D structures. In contrast, both the
chimeric 3D3 and the humanized 3D3 prevented the spheroids from forming.
The results are shown in duplicate (left and right panels). These results
indicate that the biological activity of the chimeric 3D3 was conserved
in the humanized 3D3 and suggests that the h3D3 will behave in an
identical manner.
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Sequences Referred to in the Description
TABLE-US-00006
[0568] SEQ ID NO.: 1
GAGGGGCATCAATCACACCGAGAAGTCACAGCCCCTCAACCACTGAGGTGTGGGGGGGTAGGGAT
CTGCATTTCTTCATATCAACCCCACACTATAGGGCACCTAAATGGGTGGGCGGTGGGGGAGACCG
ACTCACTTGAGTTTCTTGAAGGCTTCCTGGCCTCCAGCCACGTAATTGCCCCCGCTCTGGATCTG
GTCTAGCTTCCGGATTCGGTGGCCAGTCCGCGGGGTGTAGATGTTCCTGACGGCCCCAAAGGGTG
CCTGAACGCCGCCGGTCACCTCCTTCAGGAAGACTTCGAAGCTGGACACCTTCTTCTCATGGATG
ACGACGCGGCGCCCCGCGTAGAAGGGGTCCCCGTTGCGGTACACAAGCACGCTCTTCACGACGGG
CTGAGACAGGTGGCTGGACCTGGCGCTGCTGCCGCTCATCTTCCCCGCTGGCCGCCGCCTCAGCT
CGCTGCTTCGCGTCGGGAGGCACCTCCGCTGTCCCAGCGGCCTCACCGCACCCAGGGCGCGGGAT
CGCCTCCTGAAACGAACGAGAAACTGACGAATCCACAGGTGAAAGAGAAGTAACGGCCGTGCGCC
TAGGCGTCCACCCAGAGGAGACACTAGGAGCTTGCAGGACTCGGAGTAGACGCTCAAGTTTTTCA
CCGTGGCGTGCACAGCCAATCAGGACCCGCAGTGCGCGCACCACACCAGGTTCACCTGCTACGGG
CAGAATCAAGGTGGACAGCTTCTGAGCAGGAGCCGGAAACGCGCGGGGCCTTCAAACAGGCACGC
CTAGTGAGGGCAGGAGAGAGGAGGACGCACACACACACACACACACAAATATGGTGAAACCCAAT
TTCTTACATCATATCTGTGCTACCCTTTCCAAACAGCCTA
SEQ ID NO.: 2
MDDDAAPRVEGVPVAVHKHALHDGLRQVAGPGAAAAHLPRWPPPQLAASRREAPPLSQRPHRTQG
AGSPPETNEKLTNPQVKEK
SEQ ID NO.: 3
GACATTGTGATGACCCAGTCTCCATCCTCCCTGGCTGTGTCAATAGGACAGAAGGTCACTATGAA
CTGCAAGTCCAGTCAGAGCCTTTTAAATAGTAACTTTCAAAAGAACTTTTTGGCCTGGTACCAGC
AGAAACCAGGCCAGTCTCCTAAACTTCTGATATACTTTGCATCCACTCGGGAATCTAGTATCCCT
GATCGCTTCATAGGCAGTGGATCTGGGACAGATTTCACTCTTACCATCAGCAGTGTGCAGGCTGA
AGACCTGGCAGATTACTTCTGTCAGCAACATTATAGCACTCCGCTCACGTTCGGTGCTGGGACCA
AGCTGGAGCTGAAAGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTG
AAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACA
GTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCA
AGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAA
GTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGG
AGAGTGT
SEQ ID NO.: 4
DIVMTQSPSSLAVSIGQKVTMNCKSSQSLLNSNFQKNFLAWYQQKPGQSPKLLIYFASTRESSIP
DRFIGSGSGTDFTLTISSVQAEDLADYFCQQHYSTPLTFGAGTKLELKAVAAPSVFIFPPSDEQL
KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK
VYACEVTHQGLSSPVTKSFNRGEC
SEQ ID NO.: 5
GAGGTTCAGCTGCAGCAGTCTGTAGCTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTG
CAAGGCTTCGGGCTACATATTTACTGACTATGAGATACACTGGGTGAAGCAGACTCCTGTGCATG
GCCTGGAATGGATTGGGGTTATTGATCCTGAAACTGGTAATACTGCCTTCAATCAGAAGTTCAAG
GGCAAGGCCACACTGACTGCAGACATATCCTCCAGCACAGCCTACATGGAACTCAGCAGTTTGAC
ATCTGAGGACTCTGCCGTCTATTACTGTATGGGTTATTCTGATTATTGGGGCCAAGGCACCACTC
TCACAGTCTCCTCAGCCTCAACGAAGGGCCCATCTGTCTTTCCCCTGGCCCCCTCCTCCAAGAGC
ACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGT
GTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAG
GACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATC
TGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGA
ATTCACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCT
TCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTG
GACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAA
TGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCG
TCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA
GCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCT
GCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCT
ATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACG
CCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAG
GTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGC
AGAAGAGCCTCTCCCTGTCTCCCGGGAAA
SEQ ID NO.: 6
EVQLQQSVAELVRPGASVTLSCKASGYIFTDYEIHWVKQTPVHGLEWIGVIDPETGNTAFNQKFK
GKATLTADISSSTAYMELSSLTSEDSAVYYCMGYSDYWGQGTTLTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCEFTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVICVVV
DVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO.: 7
GATGTTTTGATGACCCAAACTCCACGCTCCCTGTCTGTCAGTCTTGGAGATCAAGCCTCCATCTC
TTGTAGATCGAGTCAGAGCCTTTTACATAGTAATGGAAACACCTATTTAGAATGGTATTTGCAGA
AACCAGGCCAGCCTCCAAAGGTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGAC
AGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCGGAGTGGAGGCTGAGGA
TCTGGGAGTTTATTACTGCTTTCAAGGTTCACATGTTCCTCTCACGTTCGGTGCTGGGACCAAGC
TGGAGCTGAAAGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTG
GAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGG
ACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTC
TACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGA
GTGT
SEQ ID NO.: 8
DVLMTQTPRSLSVSLGDQASISCRSSQSLLHSNGNTYLEWYLQKPGQPPKVLIYKVSNRFSGVPD
RFSGSGSGTDFTLKISGVEAEDLGVYYCFQGSHVPLTFGAGTKLELKAVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV
YACEVTHQGLSSPVTKSFNRGEC
SEQ ID NO.: 9
GAGATCCAGCTGCAGCAGTCTGGACCTGAGTTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTG
TAAGGCTTCTGGATACACCTTCACTGACAACTACATGAACTGGGTGAAGCAGAGCCATGGAAAGA
GCCTTGAGTGGATTGGAGATATTAATCCTTACTATGGTACTACTACCTACAACCAGAAGTTCAAG
GGCAAGGCCACATTGACTGTAGACAAGTCCTCCCGCACAGCCTACATGGAGCTCCGCGGCCTGAC
ATCTGAGGACTCTGCAGTCTATTACTGTGCAAGAGATGACTGGTTTGATTATTGGGGCCAAGGGA
CTCTGGTCACTGTCTCTGCAGCCTCAACGAAGGGCCCATCTGTCTTTCCCCTGGCCCCCTCCTCC
AAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGT
GACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGT
CCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACC
TACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATC
TTGTGAATTCACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT
TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTG
GTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGT
GCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCC
TCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCC
CTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTA
CACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAG
GCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAG
ACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAA
GAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACT
ACACGCAGAAGAGCCTCTCCCTGTCTCCCGGGAAA
SEQ ID NO.: 10
EIQLQQSGPELVKPGASVKISCKASGYTFTDNYMNWVKQSHGKSLEWIGDINPYYGTTTYNQKFK
GKATLTVDKSSRTAYMELRGLTSEDSAVYYCARDDWFDYWGQGTLVTVSAASTKGPSVFPLAPSS
KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCEFTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYK
TTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO.: 11
GACATCGTTATGTCTCAGTCTCCATCTTCCATGTATGCATCTCTAGGAGAGAGAGTCACTATCAC
TTGCAAGGCGAGTCAGGACATTCATAACTTTTTAAACTGGTTCCAGCAGAAACCAGGAAAATCTC
CAAAGACCCTGATCTTTCGTGCAAACAGATTGGTAGATGGGGTCCCATCAAGGTTCAGTGGCAGT
GGATCTGGGCAAGATTATTCTCTCACCATCAGCAGCCTGGAGTTTGAAGATTTGGGAATTTATTC
TTGTCTACAGTATGATGAGATTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAGAGCTG
TGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCT
GTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGC
CCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCC
TCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTC
ACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
SEQ ID NO.: 12
DIVMSQSPSSMYASLGERVTITCKASQDIHNFLNWFQQKPGKSPKTLIFRANRLVDGVPSRFSGS
GSGQDYSLTISSLEFEDLGIYSCLQYDEIPLTFGAGTKLELRAVAAPSVFIFPPSDEQLKSGTAS
VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEV
THQGLSSPVTKSFNRGEC
SEQ ID NO.: 13
GAGGTGCAGCTTCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTCAGTCACTTTCACTCACCTG
CACTGTCACTGGCTTCTCCATCACCAGTGGTTATGGCTGGCACTGGATCCGGCAGTTTCCAGGAA
ACAAACTGGAGTGGATGGGCTACATAAACTACGATGGTCACAATGACTACAACCCATCTCTCAAA
AGTCGAATCTCTATCACTCAAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGAC
TACTGAGGACACAGCCACATATTACTGTGCAAGCAGTTACGACGGCTTATTTGCTTACTGGGGCC
AAGGGACTCTGGTCACTGTCTCTGCAGCCTCAACGAAGGGCCCATCTGTCTTTCCCCTGGCCCCC
TCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGA
ACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCC
TACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACC
CAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCC
CAAATCTTGTGAATTCACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGT
CAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACA
TGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGT
GGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCA
GCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAAC
AAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACA
GGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGG
TCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAAC
TACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGT
GGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACA
ACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCCGGGAAA
SEQ ID NO.: 14
EVQLQESGPDLVKPSQSLSLTCTVTGFSITSGYGWHWIRQFPGNKLEWMGYINYDGHNDYNPSLK
SRISITQDTSKNQFFLQLNSVTTEDTATYYCASSYDGLFAYWGQGTLVTVSAASTKGPSVFPLAP
SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT
QTYICNVNHKPSNTKVDKKVEPKSCEFTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVT
CVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENN
YKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
SEQ ID NO.: 15
GACATTGTGATGACCCAGTCTCCATCCTCCCTGGCTGTGTCAATAGGACAGAAGGTCACTATGAA
CTGCAAGTCCAGTCAGAGCCTTTTAAATAGTAACTTTCAAAAGAACTTTTTGGCCTGGTACCAGC
AGAAACCAGGCCAGTCTCCTAAACTTCTGATATACTTTGCATCCACTCGGGAATCTAGTATCCCT
GATCGCTTCATAGGCAGTGGATCTGGGACAGATTTCACTCTTACCATCAGCAGTGTGCAGGCTGA
AGACCTGGCAGATTACTTCTGTCAGCAACATTATAGCACTCCGCTCACGTTCGGTGCTGGGACCA
AGCTGGAGCTGAAA
SEQ ID NO.: 16
DIVMTQSPSSLAVSIGQKVTMNCKSSQSLLNSNFQKNFLAWYQQKPGQSPKLLIYFASTRESSIP
DRFIGSGSGTDFTLTISSVQAEDLADYFCQQHYSTPLTFGAGTKLELK
SEQ ID NO.: 17
GAGGTTCAGCTGCAGCAGTCTGTAGCTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCCTG
CAAGGCTTCGGGCTACATATTTACTGACTATGAGATACACTGGGTGAAGCAGACTCCTGTGCATG
GCCTGGAATGGATTGGGGTTATTGATCCTGAAACTGGTAATACTGCCTTCAATCAGAAGTTCAAG
GGCAAGGCCACACTGACTGCAGACATATCCTCCAGCACAGCCTACATGGAACTCAGCAGTTTGAC
ATCTGAGGACTCTGCCGTCTATTACTGTATGGGTTATTCTGATTATTGGGGCCAAGGCACCACTC
TCACAGTCTCCTCA
SEQ ID NO.: 18
EVQLQQSVAELVRPGASVTLSCKASGYIFTDYEIHWVKQTPVHGLEWIGVIDPETGNTAFNQKFK
GKATLTADISSSTAYMELSSLTSEDSAVYYCMGYSDYWGQGTTLTVSS
SEQ ID NO.: 19
GATGTTTTGATGACCCAAACTCCACGCTCCCTGTCTGTCAGTCTTGGAGATCAAGCCTCCATCTC
TTGTAGATCGAGTCAGAGCCTTTTACATAGTAATGGAAACACCTATTTAGAATGGTATTTGCAGA
AACCAGGCCAGCCTCCAAAGGTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGAC
AGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCGGAGTGGAGGCTGAGGA
TCTGGGAGTTTATTACTGCTTTCAAGGTTCACATGTTCCTCTCACGTTCGGTGCTGGGACCAAGC
TGGAGCTGAAA
SEQ ID NO.: 20
DVLMTQTPRSLSVSLGDQASISCRSSQSLLHSNGNTYLEWYLQKPGQPPKVLIYKVSNRFSGVPD
RFSGSGSGTDFTLKISGVEAEDLGVYYCFQGSHVPLTFGAGTKLELK
SEQ ID NO.: 21
GAGATCCAGCTGCAGCAGTCTGGACCTGAGTTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTG
TAAGGCTTCTGGATACACCTTCACTGACAACTACATGAACTGGGTGAAGCAGAGCCATGGAAAGA
GCCTTGAGTGGATTGGAGATATTAATCCTTACTATGGTACTACTACCTACAACCAGAAGTTCAAG
GGCAAGGCCACATTGACTGTAGACAAGTCCTCCCGCACAGCCTACATGGAGCTCCGCGGCCTGAC
ATCTGAGGACTCTGCAGTCTATTACTGTGCAAGAGATGACTGGTTTGATTATTGGGGCCAAGGGA
CTCTGGTCACTGTCTCTGCA
SEQ ID NO.: 22
EIQLQQSGPELVKPGASVKISCKASGYTFTDNYMNWVKQSHGKSLEWIGDINPYYGTTTYNQKFK
GKATLTVDKSSRTAYMELRGLTSEDSAVYYCARDDWFDYWGQGTLVTVSA
SEQ ID NO.: 23
GACATCGTTATGTCTCAGTCTCCATCTTCCATGTATGCATCTCTAGGAGAGAGAGTCACTATCAC
TTGCAAGGCGAGTCAGGACATTCATAACTTTTTAAACTGGTTCCAGCAGAAACCAGGAAAATCTC
CAAAGACCCTGATCTTTCGTGCAAACAGATTGGTAGATGGGGTCCCATCAAGGTTCAGTGGCAGT
GGATCTGGGCAAGATTATTCTCTCACCATCAGCAGCCTGGAGTTTGAAGATTTGGGAATTTATTC
TTGTCTACAGTATGATGAGATTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAGA
SEQ ID NO.: 24
DIVMSQSPSSMYASLGERVTITCKASQDIHNFLNWFQQKPGKSPKTLIFRANRLVDGVPSRFSGS
GSGQDYSLTISSLEFEDLGIYSCLQYDEIPLTFGAGTKLELR
SEQ ID NO.: 25
GAGGTGCAGCTTCAGGAGTCAGGACCTGACCTGGTGAAACCTTCTCAGTCACTTTCACTCACCTG
CACTGTCACTGGCTTCTCCATCACCAGTGGTTATGGCTGGCACTGGATCCGGCAGTTTCCAGGAA
ACAAACTGGAGTGGATGGGCTACATAAACTACGATGGTCACAATGACTACAACCCATCTCTCAAA
AGTCGAATCTCTATCACTCAAGACACATCCAAGAACCAGTTCTTCCTGCAGTTGAATTCTGTGAC
TACTGAGGACACAGCCACATATTACTGTGCAAGCAGTTACGACGGCTTATTTGCTTACTGGGGCC
AAGGGACTCTGGTCACTGTCTCTGCA
SEQ ID NO.: 26
EVQLQESGPDLVKPSQSLSLTCTVTGFSITSGYGWHWIRQFPGNKLEWMGYINYDGHNDYNPSLK
SRISITQDTSKNQFFLQLNSVTTEDTATYYCASSYDGLFAYWGQGTLVTVSA
SEQ ID NO.: 27
KSSQSLLNSNFQKNFLA
SEQ ID NO.: 28
FASTRES
SEQ ID NO.: 29
QQHYSTPLT
SEQ ID NO.: 30
GYIFTDYEIH
SEQ ID NO.: 31
VIDPETGNTA
SEQ ID NO.: 32
MGYSDY
SEQ ID NO.: 33
RSSQSLLHSNGNTYLE
SEQ ID NO.: 34
KVSNRFS
SEQ ID NO.: 35
FQGSHVPLT
SEQ ID NO.: 36
GYTFTDNYMN
SEQ ID NO.: 37
DINPYYGTTT
SEQ ID NO.: 38
ARDDWFDY
SEQ ID NO.: 39
KASQDIHNFLN
SEQ ID NO.: 40
RANRLVD
SEQ ID NO.: 41
LQYDEIPLT
SEQ ID NO.: 42
GFSITSGYGWH
SEQ ID NO.: 43
YINYDGHND
SEQ ID NO.: 44
ASSYDGLFAY
SEQ ID NO.: 45
GAGGGGCATCAATCACACCGAGAA
SEQ ID NO.: 46
CCCCACCGCCCACCCATTTAGG
SEQ ID NO.: 47
TGAAGGTCGGAGTCAACGGATTTGGT
SEQ ID NO.: 48
CATGTGGGCCATGAGGTCCACCAC
SEQ ID NO.: 49
GGCCTCCAGCCACGTAATT
SEQ ID NO.: 50
GGCGCTGCTGCCGCTCATC
SEQ ID NO.: 51
TCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCT
TGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTG
TCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTG
AAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCCATTCGCCATTCAGGCTGCGC
AACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGGCGAAAGGGGGATG
TGCTGCAAGGCGATTAAGTTGGGTAACGCCAGGGTTTTCCCAGTCACGACGTTGTAAAACGACGG
CCAGTGCCAAGCTTTTCCAAAAAACTACCGTTGTTATAGGTGTCTCTTGAACACCTATAACAACG
GTAGTGGATCCCGCGTCCTTTCCACAAGATATATAAACCCAAGAAATCGAAATACTTTCAAGTTA
CGGTAAGCATATGATAGTCCATTTTAAAACATAATTTTAAAACTGCAAACTACCCAAGAAATTAT
TACTTTCTACGTCACGTATTTTGTACTAATATCTTTGTGTTTACAGTCAAATTAATTCTAATTAT
CTCTCTAACAGCCTTGTATCGTATATGCAAATATGAAGGAATCATGGGAAATAGGCCCTCTTCCT
GCCCGACCTTGGCGCGCGCTCGGCGCGCGGTCACGCTCCGTCACGTGGTGCGTTTTGCCTGCGCG
TCTTTCCACTGGGGAATTCATGCTTCTCCTCCCTTTAGTGAGGGTAATTCTCTCTCTCTCCCTAT
AGTGAGTCGTATTAATTCCTTCTCTTCTATAGTGTCACCTAAATCGTTGCAATTCGTAATCATGT
CATAGCTGTTTCCTGTGTGAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGC
ATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACT
GCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGA
GAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTT
CGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGA
TAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGT
TGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAG
AGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCG
CTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGG
CGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGC
TGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTC
CAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGA
GGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACA
GTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATC
CGGCAAAAAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAA
AAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAAC
TCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTA
AAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCT
TAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCC
GTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCG
AGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCA
GAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTA
AGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACG
CTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCC
CCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCC
GCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAG
ATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGA
GTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTC
ATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTC
GATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGT
GAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATA
CTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATA
CATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGC
CACCTATTGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATGCATCTCA
ATTAGTCAGCAACCAGGTGTGGAAAGTCCCCAGGCTCCCCAGCAGGCAGAAGTATGCAAAGCATG
CATCTCAATTAGTCAGCAACCATAGTCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCC
CAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCG
CCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGCTTTTGCAAA
AAGCTAGCTTGCATGCCTGCAGGTCGGCCGCCACGACCGGTGCCGCCACCATCCCCTGACCCACG
CCCCTGACCCCTCACAAGGAGACGACCTTCCATGACCGAGTACAAGCCCACGGTGCGCCTCGCCA
CCCGCGACGACGTCCCCCGGGCCGTACGCACCCTCGCCGCCGCGTTCGCCGACTACCCCGCCACG
CGCCACACCGTCGACCCGGACCGCCACATCGAGCGGGTCACCGAGCTGCAAGAACTCTTCCTCAC
GCGCGTCGGGCTCGACATCGGCAAGGTGTGGGTCGCGGACGACGGCGCCGCGGTGGCGGTCTGGA
CCACGCCGGAGAGCGTCGAAGCGGGGGCGGTGTTCGCCGAGATCGGCCCGCGCATGGCCGAGTTG
AGCGGTTCCCGGCTGGCCGCGCAGCAACAGATGGAAGGCCTCCTGGCGCCGCACCGGCCCAAGGA
GCCCGCGTGGTTCCTGGCCACCGTCGGCGTCTCGCCCGACCACCAGGGCAAGGGTCTGGGCAGCG
CCGTCGTGCTCCCCGGAGTGGAGGCGGCCGAGCGCGCCGGGGTGCCCGCCTTCCTGGAGACCTCC
GCGCCCCGCAACCTCCCCTTCTACGAGCGGCTCGGCTTCACCGTCACCGCCGACGTCGAGGTGCC
CGAAGGACCGCGCACCTGGTGCATGACCCGCAAGCCCGGTGCCTGACGCCCGCCCCACGACCCGC
AGCGCCCGACCGAAAGGAGCGCACGACCCCATGGCTCCGACCGAAGCCACCCGGGGCGGCCCCGC
CGACCCCGCACCCGCCCCCGAGGCCCACCGACTCTAGAGGATCATAATCAGCCATACCACATTTG
TAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAAT
GCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCAC
AAATTTCACAAATAAAGCATTTTTTTCACTGCAATCTAAGAAACCATTATTATCATGACATTAAC
CTATAAAAATAGGCGTATCACGAGGCCCTTTCGTC
SEQ ID NO.: 52
GTAAGCGGATCCATGGATGACGACGCGGCGCCC
SEQ ID NO.: 53
GTAAGCAAGCTTCTTCTCTTTCACCTGTGGATT
SEQ ID NO.: 54
GTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGACATTGATTATTGACTAGTTATT
AATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTT
ACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTA
TGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAA
CTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGAC
GGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTA
CATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTG
GATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTT
TGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGG
CGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCA
CTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCG
CGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGG
ACCTGAGCCAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAG
TCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGA
GGTGCTGCTGATGATGTAATTAAAGTAGGCGGTCTTGAGCCGGCGGATGGTCGAGGTGAGGTGTG
GCAGGCTTGAGATCCAGCTGTTGGGGTGAGTACTCCCTCTCAAAAGCGGGCATGACTTCTGCGCT
AAGATTGTCAGTTTCCAAAAACGAGGAGGATTTGATATTCACCTGGCCCGATCTGGCCATACACT
TGAGTGACAATGACATCCACTTTGCCTTTCTCTCCACAGGTGTCCACTCCCAGGTCCAAGTTTGC
CGCCACCATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACTG
GCGCCGGATCAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTC
TTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGT
GGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGG
TGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTC
CTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGC
CCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGT
ACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAA
GGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAA
GACCACGCCTCCCGTGTTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACA
AGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC
TACACGCAGAAGAGCCTCTCCCTGTCTCCCGGGAAAGCTAGCGGAGCCGGAAGCACAACCGAAAA
CCTGTATTTTCAGGGCGGATCCGAATTCAAGCTTGATATCTGATCCCCCGACCTCGACCTCTGGC
TAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAG
GACATATGGGAGGGCAAATCATTTGGTCGAGATCCCTCGGAGATCTCTAGCTAGAGCCCCGCCGC
CGGACGAACTAAACCTGACTACGGCATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCT
TCAGTTGGTTGGTACAACTTGCCAACTGAACCCTAAACGGGTAGCATATGCTTCCCGGGTAGTAG
TATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGCTATC
GAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATGTAGGTGGGCGGGCCAAGATAGGGGCG
CGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTT
GGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACT
ACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATC
TATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATAT
CTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGG
TAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTA
TATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATC
CTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAAT
CTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATA
TCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGG
GTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCACGATGATAAGCTGTCA
AACATGAGAATTAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGT
CATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTA
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATG
CTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTT
TTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG
AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAG
AGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGT
ATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACT
TGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGC
AGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACC
GAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAAC
CGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACA
ACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTG
GATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTG
CTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGT
AAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAG
ACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCAT
ATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTT
GATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGA
AAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAA
AACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTA
ACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCA
CTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTG
CCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAG
CGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT
GAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT
ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGG
TATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTC
AGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCT
GGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCC
TTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGA
AGC
SEQ ID NO.: 55
GTAAGCAAGCTTAGGCCGCTGGGACAGCGGAGGTGC
SEQ ID NO.: 56
GTAAGCAAGCTTGGCAGCAGCGCCAGGTCCAGC
SEQ ID NO.: 57
GTAAGCAGCGCTGTGGCTGCACCATCTGTCTTC
SEQ ID NO.: 58
GTAAGCGCTAGCCTAACACTCTCCCCTGTTGAAGC
SEQ ID NO.: 59
GCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGC
CTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATA
ACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTAC
AGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGA
AGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG
SEQ ID NO.: 60
AVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTY
SLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
SEQ ID NO.: 61
CTTGAGCCGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTAC
TCCCTCTCAAAAGCGGGCATTACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTT
GATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCT
CCACAGGTGTCCACTCCCAGGTCCAAGTTTAAACGGATCTCTAGCGAATTCATGAACTTTCTGCT
GTCTTGGGTGCATTGGAGCCTTGCCTTGCTGCTCTACCTCCACCATGCCAAGTGGTCCCAGGCTT
GAGACGGAGCTTACAGCGCTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAG
TTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGT
ACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACA
GCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACAC
AAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAG
GGGAGAGTGTTAGGGTACCGCGGCCGCTTCGAATGAGATCCCCCGACCTCGACCTCTGGCTAATA
AAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCACTCGGAAGGACAT
ATGGGAGGGCAAATCATTTGGTCGAGATCCCTCGGAGATCTCTAGCTAGAGCCCCGCCGCCGGAC
GAACTAAACCTGACTACGGCATCTCTGCCCCTTCTTCGCGGGGCAGTGCATGTAATCCCTTCAGT
TGGTTGGTACAACTTGCCAACTGGGCCCTGTTCCACATGTGACACGGGGGGGGACCAAACACAAA
GGGGTTCTCTGACTGTAGTTGACATCCTTATAAATGGATGTGCACATTTGCCAACACTGAGTGGC
TTTCATCCTGGAGCAGACTTTGCAGTCTGTGGACTGCAACACAACATTGCCTTTATGTGTAACTC
TTGGCTGAAGCTCTTACACCAATGCTGGGGGACATGTACCTCCCAGGGGCCCAGGAAGACTACGG
GAGGCTACACCAACGTCAATCAGAGGGGCCTGTGTAGCTACCGATAAGCGGACCCTCAAGAGGGC
ATTAGCAATAGTGTTTATAAGGCCCCCTTGTTAACCCTAAACGGGTAGCATATGCTTCCCGGGTA
GTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGAAGCATATGC
TATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATATCTCCCACCCCATGAGCTGTCA
CGGTTTTATTTACATGGGGTCAGGATTCCACGAGGGTAGTGAACCATTTTAGTCACAAGGGCAGT
GGCTGAAGATCAAGGAGCGGGCAGTGAACTCTCCTGAATCTTCGCCTGCTTCTTCATTCTCCTTC
GTTTAGCTAATAGAATAACTGCTGAGTTGTGAACAGTAAGGTGTATGTGAGGTGCTCGAAAACAA
GGTTTCAGGTGACGCCCCCAGAATAAAATTTGGACGGGGGGTTCAGTGGTGGCATTGTGCTATGA
CACCAATATAACCCTCACAAACCCCTTGGGCAATAAATACTAGTGTAGGAATGAAACATTCTGAA
TATCTTTAACAATAGAAATCCATGGGGTGGGGACAAGCCGTAAAGACTGGATGTCCATCTCACAC
GAATTTATGGCTATGGGCAACACATAATCCTAGTGCAATATGATACTGGGGTTATTAAGATGTGT
CCCAGGCAGGGACCAAGACAGGTGAACCATGTTGTTACACTCTATTTGTAACAAGGGGAAAGAGA
GTGGACGCCGACAGCAGCGGACTCCACTGGTTGTCTCTAACACCCCCGAAAATTAAACGGGGCTC
CACGCCAATGGGGCCCATAAACAAAGACAAGTGGCCACTCTTTTTTTTGAAATTGTGGAGTGGGG
GCACGCGTCAGCCCCCACACGCCGCCCTGCGGTTTTGGACTGTAAAATAAGGGTGTAATAACTTG
GCTGATTGTAACCCCGCTAACCACTGCGGTCAAACCACTTGCCCACAAAACCACTAATGGCACCC
CGGGGAATACCTGCATAAGTAGGTGGGCGGGCCAAGATAGGGGCGCGATTGCTGCGATCTGGAGG
ACAAATTACACACACTTGCGCCTGAGCGCCAAGCACAGGGTTGTTGGTCCTCATATTCACGAGGT
CGCTGAGAGCACGGTGGGCTAATGTTGCCATGGGTAGCATATACTACCCAAATATCTGGATAGCA
TATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGC
TATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATCC
TAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATC
TGTATCCGGGTAGCATATGCTATCCTAATAGAGATTAGGGTAGTATATGCTATCCTAATTTATAT
CTGGGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGC
ATATGCTATCCTAATCTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATG
CTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATAGGCTATC
CTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTATCCTAAT
CTGTATCCGGGTAGCATATGCTATCCTCACGATGATAAGCTGTCAAACATGAGAATTAATTCTTG
AAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTT
AGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATA
CATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAG
GAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTC
CTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGA
GTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACG
TTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCG
GGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTC
ACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAG
TGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTT
TGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATA
CCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAAC
TGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTG
CAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGT
GAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGT
TATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTG
CCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTA
AAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAAT
CCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTT
GAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTG
GTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCA
GATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCAC
CGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGT
CTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGG
TTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGC
ATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTC
GGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGG
GTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGA
AAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTC
TTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGC
TCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATAC
GCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGAC
TGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGC
TTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAG
GAAACAGCTATGACCATGATTACGCCAAGCTCTAGCTAGAGGTCGACCAATTCTCATGTTTGACA
GCTTATCATCGCAGATCCGGGCAACGTTGTTGCATTGCTGCAGGCGCAGAACTGGTAGGTATGGC
AGATCTATACATTGAATCAATATTGGCAATTAGCCATATTAGTCATTGGTTATATAGCATAAATC
AATATTGGCTATTGGCCATTGCATACGTTGTATCTATATCATAATATGTACATTTATATTGGCTC
ATGTCCAATATGACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGG
GGTCATTAGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCT
GGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCC
AATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTAC
ATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGG
CATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCAT
CGCTATTACCATGGTGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCAC
GGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGG
GACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTG
GGAGGTCTATATAAGCAGAGCTCGTTTAGTGAACCGTCAGATCCTCACTCTCTTCCGCATCGCTG
TCTGCGAGGGCCAGCTGTTGGGCTCGCGGTTGAGGACAAACTCTTCGCGGTCTTTCCAGTACTCT
TGGATCGGAAACCCGTCGGCCTCCGAACGGTACTCCGCCACCGAGGGACCTGAGCGAGTCCGCAT
CGACCGGATCGGAAAACCTCTCGAGAAAGGCGTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGC
ACCGTGGCGGGCGGCAGCGGGTGGCGGTCGGGGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTA
ATTAAAGTAGGCGGT
SEQ DI NO.: 62
ATGCCAAGTGGTCCCAGGCTGACATTGTGATGACCCAGTCTCC
SEQ ID NO.: 63
ATGCCAAGTGGTCCCAGGCTGATGTTTTGATGACCCAAACTCC
SEQ ID NO.: 64
ATGCCAAGTGGTCCCAGGCTGACATCGTTATGTCTCAGTCTCC
SEQ ID NO.: 65
GGGAAGATGAAGACAGATGGTGCAGCCACAGC
SEQ ID NO.: 66
GTAAGCGCTAGCGCCTCAACGAAGGGCCCATCTGTCTTTCCCCTGGCCCC
SEQ ID NO.: 67
GTAAGCGAATTCACAAGATTTGGGCTCAACTTTCTTG
SEQ ID NO.: 68
GCCTCCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCAC
AGCAGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAG
GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTC
AGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCA
CAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGT
SEQ ID NO.: 69
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC
SEQ ID NO.: 70
CTTGAGCCGGCGGATGGTCGAGGTGAGGTGTGGCAGGCTTGAGATCCAGCTGTTGGGGTGAGTAC
TCCCTCTCAAAAGCGGGCATTACTTCTGCGCTAAGATTGTCAGTTTCCAAAAACGAGGAGGATTT
GATATTCACCTGGCCCGATCTGGCCATACACTTGAGTGACAATGACATCCACTTTGCCTTTCTCT
CCACAGGTGTCCACTCCCAGGTCCAAGTTTGCCGCCACCATGGAGACAGACACACTCCTGCTATG
GGTACTGCTGCTCTGGGTTCCAGGTTCCACTGGCGGAGACGGAGCTTACGGGCCCATCTGTCTTT
CCCCTGGCCCCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGA
CTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCT
TCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC
AGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAA
GAAAGTTGAGCCCAAATCTTGTGAATTCACTCACACATGCCCACCGTGCCCAGCACCTGAACTCC
TGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACC
CCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTA
CGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGT
ACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGC
AAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCC
CCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCC
TGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAG
CCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAG
CAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATG
AGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCCGGGAAATGATCCCCCGAC
CTCGACCTCTGGCTAATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTC
TCTCACTCGGAAGGACATATGGGAGGGCAAATCATTTGGTCGAGATCCCTCGGAGATCTCTAGCT
AGAGCCCCGCCGCCGGACGAACTAAACCTGACTACGGCATCTCTGCCCCTTCTTCGCGGGGCAGT
GCATGTAATCCCTTCAGTTGGTTGGTACAACTTGCCAACTGAACCCTAAACGGGTAGCATATGCT
TCCCGGGTAGTAGTATATACTATCCAGACTAACCCTAATTCAATAGCATATGTTACCCAACGGGA
AGCATATGCTATCGAATTAGGGTTAGTAAAAGGGTCCTAAGGAACAGCGATGTAGGTGGGCGGGC
CAAGATAGGGGCGCGATTGCTGCGATCTGGAGGACAAATTACACACACTTGCGCCTGAGCGCCAA
GCACAGGGTTGTTGGTCCTCATATTCACGAGGTCGCTGAGAGCACGGTGGGCTAATGTTGCCATG
GGTAGCATATACTACCCAAATATCTGGATAGCATATGCTATCCTAATCTATATCTGGGTAGCATA
GGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCTA
TCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTA
ATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTAATAGA
GATTAGGGTAGTATATGCTATCCTAATTTATATCTGGGTAGCATATACTACCCAAATATCTGGAT
AGCATATGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGCAT
AGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCTAATCTATATCTGGGTAGTATATGCT
ATCCTAATTTATATCTGGGTAGCATAGGCTATCCTAATCTATATCTGGGTAGCATATGCTATCCT
AATCTATATCTGGGTAGTATATGCTATCCTAATCTGTATCCGGGTAGCATATGCTATCCTCACGA
TGATAAGCTGTCAAACATGAGAATTAATTCTTGAAGACGAAAGGGCCTCGTGATACGCCTATTTT
TATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTG
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATA
ACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCG
CCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAA
GTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG
TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGC
TATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTAT
TCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGT
AAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAA
CGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTT
GATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGC
AGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAAC
AATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCT
GGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT
GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGG
ATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGAC
CAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGT
GAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGT
CAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGC
TTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCT
TTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGT
AGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA
CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACC
GGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGA
CCTACACCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGA
AAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGG
GGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTT
TGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTC
CTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAA
CCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGT
CAGTGAGCGAGGAAGCGTACATTTATATTGGCTCATGTCCAATATGACCGCCATGTTGACATTGA
TTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTT
CCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGA
CGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTG
GAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCC
TATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACT
TTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAG
TACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT
CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCC
CGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTG
AACCGTCAGATCCTCACTCTCTTCCGCATCGCTGTCTGCGAGGGCCAGCTGTTGGGCTCGCGGTT
GAGGACAAACTCTTCGCGGTCTTTCCAGTACTCTTGGATCGGAAACCCGTCGGCCTCCGAACGGT
ACTCCGCCACCGAGGGACCTGAGCGAGTCCGCATCGACCGGATCGGAAAACCTCTCGAGAAAGGC
GTCTAACCAGTCACAGTCGCAAGGTAGGCTGAGCACCGTGGCGGGCGGCAGCGGGTGGCGGTCGG
GGTTGTTTCTGGCGGAGGTGCTGCTGATGATGTAATTAAAGTAGGCGGT
SEQ ID NO.: 71
GGGTTCCAGGTTCCACTGGCGAGGTTCAGCTGCAGCAGTCTGT
SEQ ID NO.: 72
GGGTTCCAGGTTCCACTGGCGAGGTGCAGCTTCAGGAGTCAGG
SEQ ID NO.: 73
GGGGCCAGGGGAAAGACAGATGGGCCCTTCGTTGAGGC
SEQ ID NO.: 91: Exemplary embodiment of CDRL1
K-S-S-Q-S-L-L-N/H-S/T-S/N/D-N/G-Q/N/K-K/L-N-Y-L-A
SEQ ID NO.: 92: Exemplary embodiment of CDRL1
K-A-S-Q-D-I-H-N/T-Y/F-L-N
SEQ ID NO 93: Exemplary embodiment of CDRL2
F-A-S-T-R-E-S
SEQ ID NO.: 94: Exemplary embodiment of CDRL2
L-V-S-K-L-D-S
SEQ ID NO.: 95: Exemplary embodiment of CDRL2
R-A-N-R-L-V-D
SEQ ID NO.: 96: Exemplary embodiment of CDRL3
Q-Q-H-Y-S-T-P-L-T
SEQ ID NO.: 97: Exemplary embodiment of CDRL3
W/L-Q-Y/G-D/T-A/E/H-F-P-R-T
SEQ ID NO.: 98: Exemplary embodiment of CDRH1 1
G-Y-T/I-F-T-D/E-Y-E/N-M/I/V-H
SEQ ID NO.: 99: Exemplary embodiment of CDRH1
G-F-T/S-I-T-S-G-Y-G-W-H
SEQ ID NO.: 100: Exemplary embodiment of CDRH2
V/N/G-I/L-D-P-E/A/G-T/Y-G-X-T-A
SEQ ID NO.: 101: Exemplary embodiment of CDRH2
Y-I-N/S-F/Y-N/D-G
SEQ ID NO.: 102: Exemplary embodiment of CDRH3
M-G-Y-S/A-D-Y
SEQ ID NO.: 103: Exemplary embodiment of CDRH3
A-S-S-Y-D-G-F-L-A-Y
SEQ ID NO.: 104: Exemplary embodiment of CDRH3 3
A-R/W-W/F-G-L-R-Q/N
SEQ ID NO.: 158
KSSQSLLHSDGKTYLN
SEQ ID NO.: 159
LVSKLDS
SEQ ID NO.: 160
WQGTHFPRT
SEQ ID NO.: 161
GYTFTD YNMH
SEQ ID NO.: 162
YINPYNDVTE
SEQ ID NO.: 163
AWFGL RQ
SEQ ID NO.: 164
RSSKSLLHSNGN TYLY
SEQ ID NO.: 165
RMSNLAS
SEQ ID NO.: 166
MQHLEYPYT
SEQ ID NO.: 167
GDTFTD YYMN
SEQ ID NO.: 168
DINPNYGGIT
SEQ ID NO.: 169
QAYYRNS DY
SEQ ID NO.: 170
KASQDVGTAVA
SEQ ID NO.: 171
WTSTRHT
SEQ ID NO.: 172
QQHYSIPLT
SEQ ID NO.: 173
GYIFTDYEIH
SEQ ID NO.: 174
VIDPETGNTA
SEQ ID NO.: 175
MGYSDY
SEQ ID NO.: 176
MVLQTQVFISLLLWISGAYGDIVMTQSPDSLAVSLGERATINCKSSQSLLNSNFQKNFLAWYQQK
PGQPPKLLIYFASTRESSVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYSTPLTFGQGTKL
EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK
DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
SEQ ID NO.: 177
MDWTWRILFLVAAATGTHAEVQLVQSGAEVKKPGASVKVSCKASGYIFTDYEIHWVRQAPGQGLE
WMGVIDPETGNTAFNQKFKGRVTITADTSTSTAYMELSSLTSEDTAVYYCMGYSDYWGQGTLVTV
SSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP
KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH
QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPS
DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS
LSLSPGK
SEQ ID N: 178
DIVMTQSPDSLAVSLGERATINCKSSQSLLNSNFQKNFLAWYQQKPGQPPKLLIYFASTRESSVP
DRFSGSGSGTDFTLTISSLQAEDVAVYYCQQHYSTPLTFGQGTKLEIK
SEQ ID NO.: 179
EVQLVQSGAEVKKPGASVKVSCKASGYIFTDYEIHWVRQAPGQGLEWMGVIDPETGNTAFNQKFK
GRVTITADTSTSTAYMELSSLTSEDTAVYYCMGYSDYWGQGTLVTVSS
SEQ ID NO.: 180
MVLQTQVFISLLLWISGAYGDIVMTQSPSSLSASVGDRVTITCKASQDIHNFLNWFQQKPGKAPK
TLIFRANRLVDGVPSRFSGSGSGTDYTLTISSLQPEDFATYSCLQYDEIPLTFGQGTKLEIKRTV
AAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSL
SSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
SEQ ID NO.: 181
MDWTWRILFLVAAATGTHAEVQLQESGPGLVKPSQTLSLTCTVSGFSITSGYGWHWIRQHPGKGL
EWIGYINYDGHNDYNPSLKSRVTISQDTSKNQFSLKLSSVTAADTAVYYCASSYDGLFAYWGQGT
LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS
SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVF
LFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL
TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG
FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY
TQKSLSLSPGK
SEQ ID No.: 182
DIVMTQSPSSLSASVGDRVTITCKASQDIHNFLNWFQQKPGKAPKTLIFRANRLVDGVPSRFSGS
GSGTDYTLTISSLQPEDFATYSCLQYDEIPLTFGQGTKLEIK
SEQ ID NO.: 183
EVQLQESGPGLVKPSQTLSLTCTVSGFSITSGYGWHWIRQHPGKGLEWIGYINYDGHNDYNPSLK
SRVTISQDTSKNQFSLKLSSVTAADTAVYYCASSYDGLFAYWGQGTLVTVS
TABLE-US-00007
TABLE A
Light chains variable region of selected antibodies
SEQID
NO:
3z1A02L 105 DAVMTQIPLTLSVTIGQPASLSC KSSQSLLHSDGK TYLN WLLQRPGQSPHRLIS
LVSKLDS
GVPDRFTGSGSGTDFTLKISRVEAEDLGLYYC WQGTHFPRT FAGGTNLEIK
3z1F06L 106 SIVMTQTPLTLSVTIGQPASITC KSSQSLLYSDGK TYLN WLLQRPGQSPHRLIS
LVSKLDS
GVPDGFTGSGSGTDFTLKISRVEAEDLGVYYC WQGTHFPRT FGGGTHLEIK
3z1E08L 107 DAVMTQIPLTLSVTIGQPASISC KSSQSLLHSDGK TYLN WLLQRPGQSPHRLIY
LVSKLDS
GVPDRFTGSGSGTDFTLKISRVEAEDLGVYYC WQGTHFPRT FGGGTHLEIK
3z1G10L 108 DVLMTQTPRSLSVSLGDQASISC RSSQSLLHSNGN TYLE WYLQFPGQPPKVLIY
KVSNRFS
GVPDRFSGSGSGTDFTLKISGVEAEDLGVYYC FQGSHVPLT FGAGTHLELK
3z1E10L 109 DIVMTQAAPSVPVTPGESVSISC RSSKSLLHSNGN TYLY WFLQRPGQSPQLLIY
RMSNLAS
GVPDRFSGSGSGTAFTLRISRVEAEDVGVYYC MQHLEYPYT FGGGTHLEIK
3z1A09L 110 DIVMTQSPSSLAMSLGQKVTMSC KSSQSLLNSNNQLNYLA WYQQFPGQSPKLLVY
FASTRKS
GVPDRFIGSGSGTDFTLTITSVQAEDLADYFC QQHFNTPLT FGAGTHLELK
3z1B01L 111 DIVMTQSPSSLAISVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVF
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSIPLT FGAGTHLELK
3z1G05L 112 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVF
FASTRES
GVPDRFIGSGSGTDFTLTITSVQAEDLADYFC QQHYSIPLT FGSGTHLELK
3z1B02L 113 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSIPLT FGAGTHLELK
3z1B08L 114 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1G08L 115 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1F07L 116 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLIY
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1E09L 117 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FASTRES
GVPDRFIGSGSGTEFTLTITSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1C03L 118 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FGSTRES
GVPDRFIGSGSGTDFTLTISGVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1E12L 119 DIVMTQSPSSLAMSVGQKVTMNC KSSQSLLNRSNQKNYLA WYQQFPGQSPKLLVY
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSIPLT FGAGTHLELK
4z1A02L 120 DIVMTQSPSSLAMSVGQKVTMNC KSSQSLLNNSNQKNYLA WYQQFPGQSPKLLLY
FASTRES
GVPDRFIGSGSGTYFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLDLK
3z1F10L 121 DIVMTQSPSSLTMSVGQKVTMSC KSSQSLLNTSNQLNYLA WYQQFPGQSPKLLVY
FASTTES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1F04L 122 DIVMTQSPSSLTVTAGEKVTMSC KSSQSLLNTSNQKNYLA WYQQFPGQSPKLLVY
FASTRAS
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1B11L 123 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FASTRES
GVPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1D03L 124 DIVMTQSPSSLAVSIGQKVTMNC KSSQSLLNSNFQKNFLA WYQQFPGQSPKLLIY
FASTRES
SIPDRFIGSGSGTDFTLTISSVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1C03L 125 DIVMTQSPSSLAMSVGQKVTMSC KSSQSLLNSSNQKNYLA WYQQFPGQSPKLLVY
FGSTRES
GVPDRFIGSGSGTDFTLTISGVQAEDLADYFC QQHYSTPLT FGAGTHLELK
3z1G12L 126 DIVMTQSPKFMSTSVGDRVSITC KASQDVG TAVA WYQQFPGQSPELLIY
WTSTRHT
GVPDRFSGSGSGTDFTLTISSVQAEDLADYFC QQHYSIPLT FGAGTHLELR
3z1C04L 127 DIVMSQSPSSMYASLGERVTITC KASQDIH NFLN WFQQFPGKSPKTLIF
RANRLVD
GVPSRFSGSGSGQDYSLTISSLEFEDLGIYSC LQYDEIPLT FGAGTHLELR
3z1D01L 128 DIHMTQSPSSMYASLGERVTITC KASQDIH TYLN WFQQFPGKSPETLIY
RANRLVD
GVPSRFSGSGSGQDYSLTISSLEYEDMGITYC LQYDEFPLT FGAGTHLELK
3z1C02L 129 DIQMTQSPSSMYASLGERVTLTC KASQDIH NYLN WFQQFPGKSPKTLIH
RANRLVA
GVPSRFSGSGSGQDYSLTISSLEYEDLGITYC LQYDAFPLT FGAGTHLELK
3z1E06L 130 DIQMTQSPSSMYASLGERVTLTC KASQDIH NYLN WFQQFPGKSPKTLIH
RANRLVA
GVPSRFSGSGSGQDYSLTISSLEYEDLGITYC LQYDAFPLT FGAGTHLELK
3z1H03L 131 DIVMSQSPSSMYASLGERVTITC KASQDIH RFLN WFQQFPGKSPKTLIF
HANRLVD
GVPSRFSGSGSGLDYSLTISSLEYEDMGIYFC LQYDAFPLT FGAGTHLELK
TABLE-US-00008
TABLE B
Heavy chains variable region of selected antibodies
SEQID
NO:
3z1A02H 132 HEIQLQQSGPELVKPGASVKMSCHTS GYTFTD YNMH WVKQKPGQGLEWIG
YINPYNDVTE
YNEKFKGRATLTSDKSSSTAYMDLSSLTSDDSAVYFC AWFGL RQ WGQGTLVTVST
3z1F06H 133 HEVQLQQSGPELVKPGASVKMSCKAS GYIFTE YNIH WVKQKPGQGPEWIG
NINPYNDVTE
YNEKFKGKATLTSDKASSTAYMDLSSLTSEDSAVYYC ARWGL RN WGQGTLVTVSA
3z1E08H 134 HEVQLQQSVPELVKPGASVKMSCHTS GYTFTE YNMH WVKQKPGQGPEWIG
NINPYNNVTE
YNEKFKGKATLTSDKSSSTAYLDLSSLTSEDSAVYYC ARWGL RN WGQGTLVTVSA
3z1A09H 135 HQVQVQQPGAELVRPGASVTLSCKAS GYIFTD YEVH WVRQRPVHGLEWIG
VIDPETGDTA
YNQHFKGKATLTADKSSSTAYMELSSLTAEDSAVYYC IGYA DY WGQGTTLTVSS
3z1B01H 136 HQVQLQQPGAELVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGGTA
YNQHFKGKATLTTDKSSSTAYMELRSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1B02H 137 HEVQLQQSGAELVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGATA
YNQHFKGKATLTADKSSSTAYMELSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1F04H 138 HEVQLQQSGAELVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGSTA
YNQHFKGKATLTADKASSTAYMELSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3Z1E09H 139 HEVQLQQSGAELVRPGASATLSCKAS GYTFTD YEMH WVKQTPVHGLEWIG
VIDPETGSTA
YNQHFKGKATLTADKSSSTAYMELSSLTSEDSAVYYC MGYA DY WGQGTTLTVSS
3z1B08H 140 HEVQLQQSGAELVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGDTA
YNQNFTGKATLTADKSSSTAYMELSSLTSEDSAVYYC MGYA DY WGQGTTLTVSS
3z1G08H 141 HQVQLKQSGAELVRPGASVTLSCKAS GYTFTD YEVH WVKQTPVHGLEWIG
VIDPATGDTA
YNQHFKGKATLTADKSSSTAYMEVSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1F07H 142 HQAYLQQSGAELVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGDTA
YNQHFKDKATLTADKASSTAYMELSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1E12H 143 HQVQLQQSEAELVKPGASVKLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGDTA
YNQHFKGKATLTADKSSSTAYMELSRLTSEDSAVYYC MGHS DY WGQGTTLTVSS
3Z1D03H 144 HEVQLQQSVAELVRPGASVTLSCKAS GYIFTD YEIH WVKQTPVHGLEWIG
VIDPETGNTA
FNQKFKGKATLTADKSSSTAYMELSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1G12H 145 HEVQLQQSVAELVRPGASVTVSCKAS GYIFTD YEIH WVKQTPAHGLEWIG
VIDPETGNTA
FNQKFKGKATLTADKSSSTAYMELSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1F10H 146 HEVQLQQSVAELVRPGAPVTLSCKAS GYTFTD YEVH WVKQTPVHGLEWIG
VIDPETGATA
YNQKFKGKATLTADKSSSAAYMELSRLTSEDSAVYYC MSYS DY WGQGTTLTVSS
3z1C03H 147 HEVQLQQSVAEVVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGVTA
YNQRFRDKATLTTDKSSSTAYMELSSLTSEDSAVYFC MGYS DY WGQGTTLTVSS
3z1C03H 148 HEVQLQQSVAEVVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VIDPETGVTA
YNQRFRDKATLTTDKSSSTAYMELSSLTSEDSAVYFC MGYS DY WGQGTTLTVSS
3z1G05H 149 HQVQLQQPGAELVRPGASVTLSCKAS GYTFTD YEIH WVKQTPVHGLEWIG
VLDPGTGRTA
YNQHFKDKATLSADKSSSTAYMELSSLTSEDSAVYYC MSYS DY WGPGTTLTVSS
3z1B11H 150 HEVQLQQSVAELVRPGASVTLSCKAS GYTFTD YEMH WVKQTPVRGLEWIG
VIDPATGDTA
YNQHFKGKATLTADKSSSAAFMELSSLTSEDSAVYYC MGYS DY WGQGTTLTVSS
3z1E06H 151 HQVQLQQSGAELVRPGASVTLSCKAS GYTFSD YEMH WVKQTPVHGLEWIG
GIDPETGDTV
YNQHFKGKATLTADKSSSTAYMELSSLTSEDSAVYYC ISYAM DY WGQGTSVTVSS
4z1A02H 152 HQVKLQQSGTELVRPGASVTLSCKAS GYKFTD YEMH WVKQTPVHGLEWIG
GIDPETGGTA
YNQHFKGKAILTADKSSTTAYMELRSLTSEDSAVYYC ISYAM DY WGQGTSVTVSS
3z1E10H 153 HEVQLQQSGPELVKPGASVKISCKAS GDTFTD YYMN WVIKQSHGKSLEWI
DINPNYGGIT
YNQHFKGKATLTVDTSSSTAYMELRGLTSEDSAVYYC QAYYRNS DY WGQGTTLTVSS
3Z1G10H 154 HEIQLQQSGPELVKPGASVKISCKAS GYTFTD NYMN WVIKQSHGKSLEWI
DINPYYGTTT
YNQHFKGKATLTVDKSSRTAYMELRGLTSEDSAVYYC ARDDWF DY WGQGTLVTVSA
3z1D01H 155 HEVQLQESGPDLVKPSQSLSLTCTVT GFSITSGYGWH WIRQFPGDKLEWMG YIS
FNGDYN
YNPSLKSRISITRDTSKNQFFLQLSSVTTEDTATYYC ASSYDGLFAY WGQGTLVTVSA
3z1C02H 156 HDVQLQESGPDLVKPSQSLSLTCTVT GFSITSGYGWH WIRQFPGNKLEWMG YIS
FNGDSN
YNPSLKSRISITRDTSKNQFFLQLNSVTSEDTATYYC ASSYDGLFAY WGQGPLVTVSA
3z1C04H 157 HEVQLQESGPDLVKPSQSLSLTCTVT GFSITSGYGWH WIRQFPGNKLEWMG YIN
YDGHND
YNPSLKSRISITQDTSKNQFFLQLNSVTTEDTATYYC ASSYDGLFAY WGQGTLVTVSA
Sequence CWU
1
1
3381885DNAHomo sapiensVan Den Eynde, A new antigen recognized by cytolytic
T lymphocytes on a human kidney tumor results from reverse strand
transcriptionJ. Exp. Med.190121793-18001999-12-20 1gaggggcatc aatcacaccg
agaagtcaca gcccctcaac cactgaggtg tgggggggta 60gggatctgca tttcttcata
tcaaccccac actatagggc acctaaatgg gtgggcggtg 120ggggagaccg actcacttga
gtttcttgaa ggcttcctgg cctccagcca cgtaattgcc 180cccgctctgg atctggtcta
gcttccggat tcggtggcca gtccgcgggg tgtagatgtt 240cctgacggcc ccaaagggtg
cctgaacgcc gccggtcacc tccttcagga agacttcgaa 300gctggacacc ttcttctcat
ggatgacgac gcggcgcccc gcgtagaagg ggtccccgtt 360gcggtacaca agcacgctct
tcacgacggg ctgagacagg tggctggacc tggcgctgct 420gccgctcatc ttccccgctg
gccgccgcct cagctcgctg cttcgcgtcg ggaggcacct 480ccgctgtccc agcggcctca
ccgcacccag ggcgcgggat cgcctcctga aacgaacgag 540aaactgacga atccacaggt
gaaagagaag taacggccgt gcgcctaggc gtccacccag 600aggagacact aggagcttgc
aggactcgga gtagacgctc aagtttttca ccgtggcgtg 660cacagccaat caggacccgc
agtgcgcgca ccacaccagg ttcacctgct acgggcagaa 720tcaaggtgga cagcttctga
gcaggagccg gaaacgcgcg gggccttcaa acaggcacgc 780ctagtgaggg caggagagag
gaggacgcac acacacacac acacacaaat atggtgaaac 840ccaatttctt acatcatatc
tgtgctaccc tttccaaaca gccta 885284PRTHomo sapiensVan
Den Eynde, A new antigen recognized by cytolytic T lymphocytes on a
human kidney tumor results from reverse strand transcriptionJ. Exp.
Med.190121793-18001999-12-20 2Met Asp Asp Asp Ala Ala Pro Arg Val Glu Gly
Val Pro Val Ala Val 1 5 10
15 His Lys His Ala Leu His Asp Gly Leu Arg Gln Val Ala Gly Pro Gly
20 25 30 Ala Ala
Ala Ala His Leu Pro Arg Trp Pro Pro Pro Gln Leu Ala Ala 35
40 45 Ser Arg Arg Glu Ala Pro Pro
Leu Ser Gln Arg Pro His Arg Thr Gln 50 55
60 Gly Ala Gly Ser Pro Pro Glu Thr Asn Glu Lys Leu
Thr Asn Pro Gln 65 70 75
80 Val Lys Glu Lys 3657DNAArtificial SequenceNucleotide sequence of
the 3D3 antibody light chain 3gacattgtga tgacccagtc tccatcctcc
ctggctgtgt caataggaca gaaggtcact 60atgaactgca agtccagtca gagcctttta
aatagtaact ttcaaaagaa ctttttggcc 120tggtaccagc agaaaccagg ccagtctcct
aaacttctga tatactttgc atccactcgg 180gaatctagta tccctgatcg cttcataggc
agtggatctg ggacagattt cactcttacc 240atcagcagtg tgcaggctga agacctggca
gattacttct gtcagcaaca ttatagcact 300ccgctcacgt tcggtgctgg gaccaagctg
gagctgaaag ctgtggctgc accatctgtc 360ttcatcttcc cgccatctga tgagcagttg
aaatctggaa ctgcctctgt tgtgtgcctg 420ctgaataact tctatcccag agaggccaaa
gtacagtgga aggtggataa cgccctccaa 480tcgggtaact cccaggagag tgtcacagag
caggacagca aggacagcac ctacagcctc 540agcagcaccc tgacgctgag caaagcagac
tacgagaaac acaaagtcta cgcctgcgaa 600gtcacccatc agggcctgag ctcgcccgtc
acaaagagct tcaacagggg agagtgt 6574219PRTArtificial SequenceAmino
acid sequence of the 3D3 antibody light chain 4Asp Ile Val Met Thr
Gln Ser Pro Ser Ser Leu Ala Val Ser Ile Gly 1 5
10 15 Gln Lys Val Thr Met Asn Cys Lys Ser Ser
Gln Ser Leu Leu Asn Ser 20 25
30 Asn Phe Gln Lys Asn Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly
Gln 35 40 45 Ser
Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Glu Ser Ser Ile 50
55 60 Pro Asp Arg Phe Ile Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp
Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
100 105 110 Lys Ala
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 115
120 125 Gln Leu Lys Ser Gly Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe 130 135
140 Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln 145 150 155
160 Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
165 170 175 Thr Tyr Ser
Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 180
185 190 Lys His Lys Val Tyr Ala Cys Glu
Val Thr His Gln Gly Leu Ser Ser 195 200
205 Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
215 51329DNAArtificial SequenceNucleotide
sequence of the 3D3 antibody heavy chain 5gaggttcagc tgcagcagtc
tgtagctgag ctggtgaggc ctggggcttc agtgacgctg 60tcctgcaagg cttcgggcta
catatttact gactatgaga tacactgggt gaagcagact 120cctgtgcatg gcctggaatg
gattggggtt attgatcctg aaactggtaa tactgccttc 180aatcagaagt tcaagggcaa
ggccacactg actgcagaca tatcctccag cacagcctac 240atggaactca gcagtttgac
atctgaggac tctgccgtct attactgtat gggttattct 300gattattggg gccaaggcac
cactctcaca gtctcctcag cctcaacgaa gggcccatct 360gtctttcccc tggccccctc
ctccaagagc acctctgggg gcacagcggc cctgggctgc 420ctggtcaagg actacttccc
cgaaccggtg acggtgtcgt ggaactcagg cgccctgacc 480agcggcgtgc acaccttccc
ggctgtccta cagtcctcag gactctactc cctcagcagc 540gtggtgaccg tgccctccag
cagcttgggc acccagacct acatctgcaa cgtgaatcac 600aagcccagca acaccaaggt
ggacaagaaa gttgagccca aatcttgtga attcactcac 660acatgcccac cgtgcccagc
acctgaactc ctggggggac cgtcagtctt cctcttcccc 720ccaaaaccca aggacaccct
catgatctcc cggacccctg aggtcacatg cgtggtggtg 780gacgtgagcc acgaagaccc
tgaggtcaag ttcaactggt acgtggacgg cgtggaggtg 840cataatgcca agacaaagcc
gcgggaggag cagtacaaca gcacgtaccg tgtggtcagc 900gtcctcaccg tcctgcacca
ggactggctg aatggcaagg agtacaagtg caaggtctcc 960aacaaagccc tcccagcccc
catcgagaaa accatctcca aagccaaagg gcagccccga 1020gaaccacagg tgtacaccct
gcccccatcc cgggatgagc tgaccaagaa ccaggtcagc 1080ctgacctgcc tggtcaaagg
cttctatccc agcgacatcg ccgtggagtg ggagagcaat 1140gggcagccgg agaacaacta
caagaccacg cctcccgtgc tggactccga cggctccttc 1200ttcctctaca gcaagctcac
cgtggacaag agcaggtggc agcaggggaa cgtcttctca 1260tgctccgtga tgcatgaggc
tctgcacaac cactacacgc agaagagcct ctccctgtct 1320cccgggaaa
13296443PRTArtificial
SequenceAmino acid sequence of the 3D3 antibody heavy chain 6Glu Val
Gln Leu Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly Ala 1 5
10 15 Ser Val Thr Leu Ser Cys Lys
Ala Ser Gly Tyr Ile Phe Thr Asp Tyr 20 25
30 Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly
Leu Glu Trp Ile 35 40 45
Gly Val Ile Asp Pro Glu Thr Gly Asn Thr Ala Phe Asn Gln Lys Phe
50 55 60 Lys Gly Lys
Ala Thr Leu Thr Ala Asp Ile Ser Ser Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Ser Ser Leu Thr
Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85
90 95 Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr
Thr Leu Thr Val Ser 100 105
110 Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser
Ser 115 120 125 Lys
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 130
135 140 Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr 145 150
155 160 Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr 165 170
175 Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
180 185 190 Thr Tyr
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp 195
200 205 Lys Lys Val Glu Pro Lys Ser
Cys Glu Phe Thr His Thr Cys Pro Pro 210 215
220 Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val
Phe Leu Phe Pro 225 230 235
240 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr
245 250 255 Cys Val Val
Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 260
265 270 Trp Tyr Val Asp Gly Val Glu Val
His Asn Ala Lys Thr Lys Pro Arg 275 280
285 Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
Leu Thr Val 290 295 300
Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 305
310 315 320 Asn Lys Ala Leu
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys 325
330 335 Gly Gln Pro Arg Glu Pro Gln Val Tyr
Thr Leu Pro Pro Ser Arg Asp 340 345
350 Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
Gly Phe 355 360 365
Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 370
375 380 Asn Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 385 390
395 400 Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
Ser Arg Trp Gln Gln Gly 405 410
415 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
Tyr 420 425 430 Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440
7654DNAArtificial SequenceNucleotide sequence of the 3G10
antibody light chain 7gatgttttga tgacccaaac tccacgctcc ctgtctgtca
gtcttggaga tcaagcctcc 60atctcttgta gatcgagtca gagcctttta catagtaatg
gaaacaccta tttagaatgg 120tatttgcaga aaccaggcca gcctccaaag gtcctgatct
acaaagtttc caaccgattt 180tctggggtcc cagacaggtt cagtggcagt ggatcaggga
cagatttcac actcaagatc 240agcggagtgg aggctgagga tctgggagtt tattactgct
ttcaaggttc acatgttcct 300ctcacgttcg gtgctgggac caagctggag ctgaaagctg
tggctgcacc atctgtcttc 360atcttcccgc catctgatga gcagttgaaa tctggaactg
cctctgttgt gtgcctgctg 420aataacttct atcccagaga ggccaaagta cagtggaagg
tggataacgc cctccaatcg 480ggtaactccc aggagagtgt cacagagcag gacagcaagg
acagcaccta cagcctcagc 540agcaccctga cgctgagcaa agcagactac gagaaacaca
aagtctacgc ctgcgaagtc 600acccatcagg gcctgagctc gcccgtcaca aagagcttca
acaggggaga gtgt 6548218PRTArtificial SequenceAmino acid sequence
of the 3G10 antibody light chain 8Asp Val Leu Met Thr Gln Thr Pro
Arg Ser Leu Ser Val Ser Leu Gly 1 5 10
15 Asp Gln Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
Leu His Ser 20 25 30
Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly Gln Pro
35 40 45 Pro Lys Val Leu
Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50
55 60 Asp Arg Phe Ser Gly Ser Gly Ser
Gly Thr Asp Phe Thr Leu Lys Ile 65 70
75 80 Ser Gly Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr
Cys Phe Gln Gly 85 90
95 Ser His Val Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys
100 105 110 Ala Val Ala
Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 115
120 125 Leu Lys Ser Gly Thr Ala Ser Val
Val Cys Leu Leu Asn Asn Phe Tyr 130 135
140 Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala
Leu Gln Ser 145 150 155
160 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
165 170 175 Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 180
185 190 His Lys Val Tyr Ala Cys Glu Val Thr
His Gln Gly Leu Ser Ser Pro 195 200
205 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 210
215 91335DNAArtificial SequenceNucleotide sequence of
the 3G10 antibody heavy chain 9gagatccagc tgcagcagtc tggacctgag
ttggtgaagc ctggggcttc agtgaagata 60tcctgtaagg cttctggata caccttcact
gacaactaca tgaactgggt gaagcagagc 120catggaaaga gccttgagtg gattggagat
attaatcctt actatggtac tactacctac 180aaccagaagt tcaagggcaa ggccacattg
actgtagaca agtcctcccg cacagcctac 240atggagctcc gcggcctgac atctgaggac
tctgcagtct attactgtgc aagagatgac 300tggtttgatt attggggcca agggactctg
gtcactgtct ctgcagcctc aacgaagggc 360ccatctgtct ttcccctggc cccctcctcc
aagagcacct ctgggggcac agcggccctg 420ggctgcctgg tcaaggacta cttccccgaa
ccggtgacgg tgtcgtggaa ctcaggcgcc 480ctgaccagcg gcgtgcacac cttcccggct
gtcctacagt cctcaggact ctactccctc 540agcagcgtgg tgaccgtgcc ctccagcagc
ttgggcaccc agacctacat ctgcaacgtg 600aatcacaagc ccagcaacac caaggtggac
aagaaagttg agcccaaatc ttgtgaattc 660actcacacat gcccaccgtg cccagcacct
gaactcctgg ggggaccgtc agtcttcctc 720ttccccccaa aacccaagga caccctcatg
atctcccgga cccctgaggt cacatgcgtg 780gtggtggacg tgagccacga agaccctgag
gtcaagttca actggtacgt ggacggcgtg 840gaggtgcata atgccaagac aaagccgcgg
gaggagcagt acaacagcac gtaccgtgtg 900gtcagcgtcc tcaccgtcct gcaccaggac
tggctgaatg gcaaggagta caagtgcaag 960gtctccaaca aagccctccc agcccccatc
gagaaaacca tctccaaagc caaagggcag 1020ccccgagaac cacaggtgta caccctgccc
ccatcccggg atgagctgac caagaaccag 1080gtcagcctga cctgcctggt caaaggcttc
tatcccagcg acatcgccgt ggagtgggag 1140agcaatgggc agccggagaa caactacaag
accacgcctc ccgtgctgga ctccgacggc 1200tccttcttcc tctacagcaa gctcaccgtg
gacaagagca ggtggcagca ggggaacgtc 1260ttctcatgct ccgtgatgca tgaggctctg
cacaaccact acacgcagaa gagcctctcc 1320ctgtctcccg ggaaa
133510445PRTArtificial SequenceAmino
acid sequence of the 3G10 antibody heavy chain 10Glu Ile Gln Leu Gln
Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5
10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly
Tyr Thr Phe Thr Asp Asn 20 25
30 Tyr Met Asn Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp
Ile 35 40 45 Gly
Asp Ile Asn Pro Tyr Tyr Gly Thr Thr Thr Tyr Asn Gln Lys Phe 50
55 60 Lys Gly Lys Ala Thr Leu
Thr Val Asp Lys Ser Ser Arg Thr Ala Tyr 65 70
75 80 Met Glu Leu Arg Gly Leu Thr Ser Glu Asp Ser
Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Asp Asp Trp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110 Val Ser
Ala Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 115
120 125 Ser Ser Lys Ser Thr Ser Gly
Gly Thr Ala Ala Leu Gly Cys Leu Val 130 135
140 Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn Ser Gly Ala 145 150 155
160 Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
165 170 175 Leu Tyr Ser
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 180
185 190 Thr Gln Thr Tyr Ile Cys Asn Val
Asn His Lys Pro Ser Asn Thr Lys 195 200
205 Val Asp Lys Lys Val Glu Pro Lys Ser Cys Glu Phe Thr
His Thr Cys 210 215 220
Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 225
230 235 240 Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 245
250 255 Val Thr Cys Val Val Val Asp Val Ser
His Glu Asp Pro Glu Val Lys 260 265
270 Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
Thr Lys 275 280 285
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 290
295 300 Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 305 310
315 320 Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu Lys Thr Ile Ser Lys 325 330
335 Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
Ser 340 345 350 Arg
Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 355
360 365 Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 370 375
380 Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp Ser Asp Gly 385 390 395
400 Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
405 410 415 Gln Gly
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 420
425 430 His Tyr Thr Gln Lys Ser Leu
Ser Leu Ser Pro Gly Lys 435 440
445 11639DNAArtificial SequenceNucleotide sequence of the 3C4 antibody
light chain 11gacatcgtta tgtctcagtc tccatcttcc atgtatgcat ctctaggaga
gagagtcact 60atcacttgca aggcgagtca ggacattcat aactttttaa actggttcca
gcagaaacca 120ggaaaatctc caaagaccct gatctttcgt gcaaacagat tggtagatgg
ggtcccatca 180aggttcagtg gcagtggatc tgggcaagat tattctctca ccatcagcag
cctggagttt 240gaagatttgg gaatttattc ttgtctacag tatgatgaga ttccgctcac
gttcggtgct 300gggaccaagc tggagctgag agctgtggct gcaccatctg tcttcatctt
cccgccatct 360gatgagcagt tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa
cttctatccc 420agagaggcca aagtacagtg gaaggtggat aacgccctcc aatcgggtaa
ctcccaggag 480agtgtcacag agcaggacag caaggacagc acctacagcc tcagcagcac
cctgacgctg 540agcaaagcag actacgagaa acacaaagtc tacgcctgcg aagtcaccca
tcagggcctg 600agctcgcccg tcacaaagag cttcaacagg ggagagtgt
63912213PRTArtificial SequenceAmino acid sequence of the 3C4
antibody light chain 12Asp Ile Val Met Ser Gln Ser Pro Ser Ser Met
Tyr Ala Ser Leu Gly 1 5 10
15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile His Asn Phe
20 25 30 Leu Asn
Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35
40 45 Phe Arg Ala Asn Arg Leu Val
Asp Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser
Ser Leu Glu Phe 65 70 75
80 Glu Asp Leu Gly Ile Tyr Ser Cys Leu Gln Tyr Asp Glu Ile Pro Leu
85 90 95 Thr Phe Gly
Ala Gly Thr Lys Leu Glu Leu Arg Ala Val Ala Ala Pro 100
105 110 Ser Val Phe Ile Phe Pro Pro Ser
Asp Glu Gln Leu Lys Ser Gly Thr 115 120
125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg
Glu Ala Lys 130 135 140
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145
150 155 160 Ser Val Thr Glu
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165
170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr
Glu Lys His Lys Val Tyr Ala 180 185
190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
Ser Phe 195 200 205
Asn Arg Gly Glu Cys 210 131341DNAArtificial
SequenceNucleotide sequence of the 3C4 antibody heavy chain
13gaggtgcagc ttcaggagtc aggacctgac ctggtgaaac cttctcagtc actttcactc
60acctgcactg tcactggctt ctccatcacc agtggttatg gctggcactg gatccggcag
120tttccaggaa acaaactgga gtggatgggc tacataaact acgatggtca caatgactac
180aacccatctc tcaaaagtcg aatctctatc actcaagaca catccaagaa ccagttcttc
240ctgcagttga attctgtgac tactgaggac acagccacat attactgtgc aagcagttac
300gacggcttat ttgcttactg gggccaaggg actctggtca ctgtctctgc agcctcaacg
360aagggcccat ctgtctttcc cctggccccc tcctccaaga gcacctctgg gggcacagcg
420gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca
480ggcgccctga ccagcggcgt gcacaccttc ccggctgtcc tacagtcctc aggactctac
540tccctcagca gcgtggtgac cgtgccctcc agcagcttgg gcacccagac ctacatctgc
600aacgtgaatc acaagcccag caacaccaag gtggacaaga aagttgagcc caaatcttgt
660gaattcactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc
720ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca
780tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac
840ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac
900cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag
960tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa
1020gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggatga gctgaccaag
1080aaccaggtca gcctgacctg cctggtcaaa ggcttctatc ccagcgacat cgccgtggag
1140tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc
1200gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg gcagcagggg
1260aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc
1320ctctccctgt ctcccgggaa a
134114447PRTArtificial SequenceAmino acid sequence of the 3C4 antibody
heavy chain 14Glu Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys
Pro Ser Gln 1 5 10 15
Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Phe Ser Ile Thr Ser Gly
20 25 30 Tyr Gly Trp His
Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp 35
40 45 Met Gly Tyr Ile Asn Tyr Asp Gly His
Asn Asp Tyr Asn Pro Ser Leu 50 55
60 Lys Ser Arg Ile Ser Ile Thr Gln Asp Thr Ser Lys Asn
Gln Phe Phe 65 70 75
80 Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys
85 90 95 Ala Ser Ser Tyr
Asp Gly Leu Phe Ala Tyr Trp Gly Gln Gly Thr Leu 100
105 110 Val Thr Val Ser Ala Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro Leu 115 120
125 Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
Gly Cys 130 135 140
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser 145
150 155 160 Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165
170 175 Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
Thr Val Pro Ser Ser Ser 180 185
190 Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser
Asn 195 200 205 Thr
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Glu Phe Thr His 210
215 220 Thr Cys Pro Pro Cys Pro
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val 225 230
235 240 Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr 245 250
255 Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu
260 265 270 Val Lys
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275
280 285 Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295
300 Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys 305 310 315
320 Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335 Ser Lys Ala
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340
345 350 Pro Ser Arg Asp Glu Leu Thr Lys
Asn Gln Val Ser Leu Thr Cys Leu 355 360
365 Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
Glu Ser Asn 370 375 380
Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 385
390 395 400 Asp Gly Ser Phe
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405
410 415 Trp Gln Gln Gly Asn Val Phe Ser Cys
Ser Val Met His Glu Ala Leu 420 425
430 His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
Lys 435 440 445
15339DNAArtificial SequenceNucleotide sequence of the 3D3 antibody light
chain variable region 15gacattgtga tgacccagtc tccatcctcc ctggctgtgt
caataggaca gaaggtcact 60atgaactgca agtccagtca gagcctttta aatagtaact
ttcaaaagaa ctttttggcc 120tggtaccagc agaaaccagg ccagtctcct aaacttctga
tatactttgc atccactcgg 180gaatctagta tccctgatcg cttcataggc agtggatctg
ggacagattt cactcttacc 240atcagcagtg tgcaggctga agacctggca gattacttct
gtcagcaaca ttatagcact 300ccgctcacgt tcggtgctgg gaccaagctg gagctgaaa
33916113PRTArtificial SequenceAmino acid sequence
of the 3D3 antibody light chain variable region 16Asp Ile Val Met
Thr Gln Ser Pro Ser Ser Leu Ala Val Ser Ile Gly 1 5
10 15 Gln Lys Val Thr Met Asn Cys Lys Ser
Ser Gln Ser Leu Leu Asn Ser 20 25
30 Asn Phe Gln Lys Asn Phe Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45
Ser Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Glu Ser Ser Ile 50
55 60 Pro Asp Arg Phe Ile
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
Asp Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu
Leu 100 105 110 Lys
17339DNAArtificial SequenceNucleotide sequence of the 3D3 antibody heavy
chain variable region 17gaggttcagc tgcagcagtc tgtagctgag ctggtgaggc
ctggggcttc agtgacgctg 60tcctgcaagg cttcgggcta catatttact gactatgaga
tacactgggt gaagcagact 120cctgtgcatg gcctggaatg gattggggtt attgatcctg
aaactggtaa tactgccttc 180aatcagaagt tcaagggcaa ggccacactg actgcagaca
tatcctccag cacagcctac 240atggaactca gcagtttgac atctgaggac tctgccgtct
attactgtat gggttattct 300gattattggg gccaaggcac cactctcaca gtctcctca
33918113PRTArtificial SequenceAmino acid sequence
of the 3D3 antibody heavy chain variable region 18Glu Val Gln Leu
Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly Ala 1 5
10 15 Ser Val Thr Leu Ser Cys Lys Ala Ser
Gly Tyr Ile Phe Thr Asp Tyr 20 25
30 Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu
Trp Ile 35 40 45
Gly Val Ile Asp Pro Glu Thr Gly Asn Thr Ala Phe Asn Gln Lys Phe 50
55 60 Lys Gly Lys Ala Thr
Leu Thr Ala Asp Ile Ser Ser Ser Thr Ala Tyr 65 70
75 80 Met Glu Leu Ser Ser Leu Thr Ser Glu Asp
Ser Ala Val Tyr Tyr Cys 85 90
95 Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val
Ser 100 105 110 Ser
19336DNAArtificial SequenceNucleotide sequence of the 3G10 antibody light
chain variable region 19gatgttttga tgacccaaac tccacgctcc ctgtctgtca
gtcttggaga tcaagcctcc 60atctcttgta gatcgagtca gagcctttta catagtaatg
gaaacaccta tttagaatgg 120tatttgcaga aaccaggcca gcctccaaag gtcctgatct
acaaagtttc caaccgattt 180tctggggtcc cagacaggtt cagtggcagt ggatcaggga
cagatttcac actcaagatc 240agcggagtgg aggctgagga tctgggagtt tattactgct
ttcaaggttc acatgttcct 300ctcacgttcg gtgctgggac caagctggag ctgaaa
33620112PRTArtificial SequenceAmino acid sequence
of the 3G10 antibody light chain variable region 20Asp Val Leu Met
Thr Gln Thr Pro Arg Ser Leu Ser Val Ser Leu Gly 1 5
10 15 Asp Gln Ala Ser Ile Ser Cys Arg Ser
Ser Gln Ser Leu Leu His Ser 20 25
30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr Leu Gln Lys Pro Gly
Gln Pro 35 40 45
Pro Lys Val Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro 50
55 60 Asp Arg Phe Ser Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70
75 80 Ser Gly Val Glu Ala Glu Asp Leu Gly Val
Tyr Tyr Cys Phe Gln Gly 85 90
95 Ser His Val Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
Lys 100 105 110
21345DNAArtificial SequenceNucleotide sequence of the 3G10 antibody heavy
chain variable region 21gagatccagc tgcagcagtc tggacctgag ttggtgaagc
ctggggcttc agtgaagata 60tcctgtaagg cttctggata caccttcact gacaactaca
tgaactgggt gaagcagagc 120catggaaaga gccttgagtg gattggagat attaatcctt
actatggtac tactacctac 180aaccagaagt tcaagggcaa ggccacattg actgtagaca
agtcctcccg cacagcctac 240atggagctcc gcggcctgac atctgaggac tctgcagtct
attactgtgc aagagatgac 300tggtttgatt attggggcca agggactctg gtcactgtct
ctgca 34522115PRTArtificial SequenceAmino acid
sequence of the 3G10 antibody heavy chain variable region 22Glu Ile
Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala 1 5
10 15 Ser Val Lys Ile Ser Cys Lys
Ala Ser Gly Tyr Thr Phe Thr Asp Asn 20 25
30 Tyr Met Asn Trp Val Lys Gln Ser His Gly Lys Ser
Leu Glu Trp Ile 35 40 45
Gly Asp Ile Asn Pro Tyr Tyr Gly Thr Thr Thr Tyr Asn Gln Lys Phe
50 55 60 Lys Gly Lys
Ala Thr Leu Thr Val Asp Lys Ser Ser Arg Thr Ala Tyr 65
70 75 80 Met Glu Leu Arg Gly Leu Thr
Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Asp Asp Trp Phe Asp Tyr Trp Gly Gln
Gly Thr Leu Val Thr 100 105
110 Val Ser Ala 115 23321DNAArtificial SequenceNucleotide
sequence of the 3C4 antibody light chain variable region
23gacatcgtta tgtctcagtc tccatcttcc atgtatgcat ctctaggaga gagagtcact
60atcacttgca aggcgagtca ggacattcat aactttttaa actggttcca gcagaaacca
120ggaaaatctc caaagaccct gatctttcgt gcaaacagat tggtagatgg ggtcccatca
180aggttcagtg gcagtggatc tgggcaagat tattctctca ccatcagcag cctggagttt
240gaagatttgg gaatttattc ttgtctacag tatgatgaga ttccgctcac gttcggtgct
300gggaccaagc tggagctgag a
32124107PRTArtificial SequenceAmino acid sequence of the 3C4 antibody
light chain variable region 24Asp Ile Val Met Ser Gln Ser Pro Ser
Ser Met Tyr Ala Ser Leu Gly 1 5 10
15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile His
Asn Phe 20 25 30
Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile
35 40 45 Phe Arg Ala Asn
Arg Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50
55 60 Ser Gly Ser Gly Gln Asp Tyr Ser
Leu Thr Ile Ser Ser Leu Glu Phe 65 70
75 80 Glu Asp Leu Gly Ile Tyr Ser Cys Leu Gln Tyr Asp
Glu Ile Pro Leu 85 90
95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Arg 100
105 25351DNAArtificial SequenceNucleotide sequence of
the 3C4 antibody heavy chain variable region 25gaggtgcagc ttcaggagtc
aggacctgac ctggtgaaac cttctcagtc actttcactc 60acctgcactg tcactggctt
ctccatcacc agtggttatg gctggcactg gatccggcag 120tttccaggaa acaaactgga
gtggatgggc tacataaact acgatggtca caatgactac 180aacccatctc tcaaaagtcg
aatctctatc actcaagaca catccaagaa ccagttcttc 240ctgcagttga attctgtgac
tactgaggac acagccacat attactgtgc aagcagttac 300gacggcttat ttgcttactg
gggccaaggg actctggtca ctgtctctgc a 35126117PRTArtificial
SequenceAmino acid sequence of the 3C4 antibody heavy chain variable
region 26Glu Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln
1 5 10 15 Ser Leu
Ser Leu Thr Cys Thr Val Thr Gly Phe Ser Ile Thr Ser Gly 20
25 30 Tyr Gly Trp His Trp Ile Arg
Gln Phe Pro Gly Asn Lys Leu Glu Trp 35 40
45 Met Gly Tyr Ile Asn Tyr Asp Gly His Asn Asp Tyr
Asn Pro Ser Leu 50 55 60
Lys Ser Arg Ile Ser Ile Thr Gln Asp Thr Ser Lys Asn Gln Phe Phe 65
70 75 80 Leu Gln Leu
Asn Ser Val Thr Thr Glu Asp Thr Ala Thr Tyr Tyr Cys 85
90 95 Ala Ser Ser Tyr Asp Gly Leu Phe
Ala Tyr Trp Gly Gln Gly Thr Leu 100 105
110 Val Thr Val Ser Ala 115
2717PRTArtificial SequenceAmino acid sequence of the 3D3 light chain CDR1
27Lys Ser Ser Gln Ser Leu Leu Asn Ser Asn Phe Gln Lys Asn Phe Leu 1
5 10 15 Ala
287PRTArtificial SequenceAmino acid sequence of the 3D3 light chain CDR2
28Phe Ala Ser Thr Arg Glu Ser 1 5
299PRTArtificial SequenceAmino acid sequence of the 3D3 light chain CDR3
29Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
3010PRTArtificial SequenceAmino acid sequence of the 3D3 heavy chain CDR1
30Gly Tyr Ile Phe Thr Asp Tyr Glu Ile His 1 5
10 3110PRTArtificial SequenceAmino acid sequence of the 3D3 heavy
chain CDR2 31Val Ile Asp Pro Glu Thr Gly Asn Thr Ala 1 5
10 326PRTArtificial SequenceAmino acid sequence of the
3D3 heavy chain CDR3 32Met Gly Tyr Ser Asp Tyr 1 5
3316PRTArtificial SequenceAmino acid sequence of the 3G10 light chain
CDR1 33Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Asn Thr Tyr Leu Glu 1
5 10 15
347PRTArtificial SequenceAmino acid sequence of the 3G10 light chain
CDR2 34Lys Val Ser Asn Arg Phe Ser 1 5
359PRTArtificial SequenceAmino acid sequence of the 3G10 light chain
CDR3 35Phe Gln Gly Ser His Val Pro Leu Thr 1 5
3610PRTArtificial SequenceAmino acid sequence of the 3G10 heavy
chain CDR1 36Gly Tyr Thr Phe Thr Asp Asn Tyr Met Asn 1
5 10 3710PRTArtificial SequenceAmino acid sequence of
the 3G10 heavy chain CDR2 37Asp Ile Asn Pro Tyr Tyr Gly Thr Thr Thr
1 5 10 388PRTArtificial SequenceAmino
acid sequence of the 3G10 heavy chain CDR3 38Ala Arg Asp Asp Trp Phe
Asp Tyr 1 5 3911PRTArtificial SequenceAmino
acid sequence of the 3C4 light chain CDR1 39Lys Ala Ser Gln Asp Ile His
Asn Phe Leu Asn 1 5 10
407PRTArtificial SequenceAmino acid sequence of the 3C4 light chain CDR2
40Arg Ala Asn Arg Leu Val Asp 1 5
419PRTArtificial SequenceAmino acid sequence of the 3C4 light chain CDR3
41Leu Gln Tyr Asp Glu Ile Pro Leu Thr 1 5
4211PRTArtificial SequenceAmino acid sequence of the 3C4 heavy chain CDR1
42Gly Phe Ser Ile Thr Ser Gly Tyr Gly Trp His 1 5
10 439PRTArtificial SequenceAmino acid sequence of the 3C4
heavy chain CDR2 43Tyr Ile Asn Tyr Asp Gly His Asn Asp 1 5
4410PRTArtificial SequenceAmino acid sequence of the 3C4
heavy chain CDR3 44Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr 1
5 10 4524DNAArtificial SequencePrimer to amplify
KAAG1 mRNA sequence 45gaggggcatc aatcacaccg agaa
244622DNAArtificial SequencePrimer to amplify KAAG1
mRNA sequence 46ccccaccgcc cacccattta gg
224726DNAArtificial SequencePrimer to amplify GAPDH gene
47tgaaggtcgg agtcaacgga tttggt
264824DNAArtificial SequencePrimer to amplify GAPDH gene 48catgtgggcc
atgaggtcca ccac
244919DNAArtificial Sequence19-mer used to generate KAAG1-specific shRNA
49ggcctccagc cacgtaatt
195019DNAArtificial Sequence19-mer used to generate KAAG1-specific shRNA
50ggcgctgctg ccgctcatc
19514455DNAArtificial SequencepSilencer 2.0 plasmid 51tcgcgcgttt
cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 60cagcttgtct
gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 120ttggcgggtg
tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 180accatatgcg
gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc 240attcgccatt
caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 300tacgccagct
ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 360tttcccagtc
acgacgttgt aaaacgacgg ccagtgccaa gcttttccaa aaaactaccg 420ttgttatagg
tgtctcttga acacctataa caacggtagt ggatcccgcg tcctttccac 480aagatatata
aacccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc 540attttaaaac
ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac 600gtattttgta
ctaatatctt tgtgtttaca gtcaaattaa ttctaattat ctctctaaca 660gccttgtatc
gtatatgcaa atatgaagga atcatgggaa ataggccctc ttcctgcccg 720accttggcgc
gcgctcggcg cgcggtcacg ctccgtcacg tggtgcgttt tgcctgcgcg 780tctttccact
ggggaattca tgcttctcct ccctttagtg agggtaattc tctctctctc 840cctatagtga
gtcgtattaa ttccttctct tctatagtgt cacctaaatc gttgcaattc 900gtaatcatgt
catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 960atacgagccg
gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 1020ttaattgcgt
tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 1080taatgaatcg
gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 1140tcgctcactg
actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 1200aaggcggtaa
tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 1260aaaggccagc
aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 1320ctccgccccc
ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 1380acaggactat
aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 1440ccgaccctgc
cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 1500tctcatagct
cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 1560tgtgtgcacg
aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 1620gagtccaacc
cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 1680agcagagcga
ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 1740tacactagaa
gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 1800agagttggta
gctcttgatc cggcaaaaaa accaccgctg gtagcggtgg tttttttgtt 1860tgcaagcagc
agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 1920acggggtctg
acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 1980tcaaaaagga
tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 2040agtatatatg
agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 2100tcagcgatct
gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 2160acgatacggg
agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 2220tcaccggctc
cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 2280ggtcctgcaa
ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 2340agtagttcgc
cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 2400tcacgctcgt
cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 2460acatgatccc
ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 2520agaagtaagt
tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 2580actgtcatgc
catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 2640tgagaatagt
gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 2700gcgccacata
gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 2760ctctcaagga
tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 2820tgatcttcag
catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 2880aatgccgcaa
aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 2940tttcaatatt
attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 3000tgtatttaga
aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 3060attggtgtgg
aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3120attagtcagc
aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3180gcatgcatct
caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3240taactccgcc
cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3300cagaggccga
ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 3360gaggcctagg
cttttgcaaa aagctagctt gcatgcctgc aggtcggccg ccacgaccgg 3420tgccgccacc
atcccctgac ccacgcccct gacccctcac aaggagacga ccttccatga 3480ccgagtacaa
gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 3540ccctcgccgc
cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 3600acatcgagcg
ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 3660gcaaggtgtg
ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 3720tcgaagcggg
ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 3780ggctggccgc
gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 3840cgtggttcct
ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 3900ccgtcgtgct
ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 3960cctccgcgcc
ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 4020tcgaggtgcc
cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc 4080cgccccacga
cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc tccgaccgaa 4140gccacccggg
gcggccccgc cgaccccgca cccgcccccg aggcccaccg actctagagg 4200atcataatca
gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 4260ctccccctga
acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 4320gcttataatg
gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 4380tcactgcaat
ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca 4440cgaggccctt
tcgtc
44555233DNAArtificial Sequenceforward primer containing BamHI site to
amplify KAAG1 cDNA 52gtaagcggat ccatggatga cgacgcggcg ccc
335333DNAArtificial Sequencereverse primer
containing HindIII site to amplify KAAG1 cDNA 53gtaagcaagc
ttcttctctt tcacctgtgg att
33545138DNAArtificial SequencepYD5 vector 54gtacatttat attggctcat
gtccaatatg accgccatgt tgacattgat tattgactag 60ttattaatag taatcaatta
cggggtcatt agttcatagc ccatatatgg agttccgcgt 120tacataactt acggtaaatg
gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 180gtcaataatg acgtatgttc
ccatagtaac gccaataggg actttccatt gacgtcaatg 240ggtggagtat ttacggtaaa
ctgcccactt ggcagtacat caagtgtatc atatgccaag 300tccgccccct attgacgtca
atgacggtaa atggcccgcc tggcattatg cccagtacat 360gaccttacgg gactttccta
cttggcagta catctacgta ttagtcatcg ctattaccat 420ggtgatgcgg ttttggcagt
acaccaatgg gcgtggatag cggtttgact cacggggatt 480tccaagtctc caccccattg
acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 540ctttccaaaa tgtcgtaata
accccgcccc gttgacgcaa atgggcggta ggcgtgtacg 600gtgggaggtc tatataagca
gagctcgttt agtgaaccgt cagatcctca ctctcttccg 660catcgctgtc tgcgagggcc
agctgttggg ctcgcggttg aggacaaact cttcgcggtc 720tttccagtac tcttggatcg
gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg 780acctgagcca gtccgcatcg
accggatcgg aaaacctctc gagaaaggcg tctaaccagt 840cacagtcgca aggtaggctg
agcaccgtgg cgggcggcag cgggtggcgg tcggggttgt 900ttctggcgga ggtgctgctg
atgatgtaat taaagtaggc ggtcttgagc cggcggatgg 960tcgaggtgag gtgtggcagg
cttgagatcc agctgttggg gtgagtactc cctctcaaaa 1020gcgggcatga cttctgcgct
aagattgtca gtttccaaaa acgaggagga tttgatattc 1080acctggcccg atctggccat
acacttgagt gacaatgaca tccactttgc ctttctctcc 1140acaggtgtcc actcccaggt
ccaagtttgc cgccaccatg gagacagaca cactcctgct 1200atgggtactg ctgctctggg
ttccaggttc cactggcgcc ggatcaactc acacatgccc 1260accgtgccca gcacctgaac
tcctgggggg accgtcagtc ttcctcttcc ccccaaaacc 1320caaggacacc ctcatgatct
cccggacccc tgaggtcaca tgcgtggtgg tggacgtgag 1380ccacgaagac cctgaggtca
agttcaactg gtacgtggac ggcgtggagg tgcataatgc 1440caagacaaag ccgcgggagg
agcagtacaa cagcacgtac cgtgtggtca gcgtcctcac 1500cgtcctgcac caggactggc
tgaatggcaa ggagtacaag tgcaaggtct ccaacaaagc 1560cctcccagcc cccatcgaga
aaaccatctc caaagccaaa gggcagcccc gagaaccaca 1620ggtgtacacc ctgcccccat
cccgggatga gctgaccaag aaccaggtca gcctgacctg 1680cctggtcaaa ggcttctatc
ccagcgacat cgccgtggag tgggagagca atgggcagcc 1740ggagaacaac tacaagacca
cgcctcccgt gttggactcc gacggctcct tcttcctcta 1800cagcaagctc accgtggaca
agagcaggtg gcagcagggg aacgtcttct catgctccgt 1860gatgcatgag gctctgcaca
accactacac gcagaagagc ctctccctgt ctcccgggaa 1920agctagcgga gccggaagca
caaccgaaaa cctgtatttt cagggcggat ccgaattcaa 1980gcttgatatc tgatcccccg
acctcgacct ctggctaata aaggaaattt attttcattg 2040caatagtgtg ttggaatttt
ttgtgtctct cactcggaag gacatatggg agggcaaatc 2100atttggtcga gatccctcgg
agatctctag ctagagcccc gccgccggac gaactaaacc 2160tgactacggc atctctgccc
cttcttcgcg gggcagtgca tgtaatccct tcagttggtt 2220ggtacaactt gccaactgaa
ccctaaacgg gtagcatatg cttcccgggt agtagtatat 2280actatccaga ctaaccctaa
ttcaatagca tatgttaccc aacgggaagc atatgctatc 2340gaattagggt tagtaaaagg
gtcctaagga acagcgatgt aggtgggcgg gccaagatag 2400gggcgcgatt gctgcgatct
ggaggacaaa ttacacacac ttgcgcctga gcgccaagca 2460cagggttgtt ggtcctcata
ttcacgaggt cgctgagagc acggtgggct aatgttgcca 2520tgggtagcat atactaccca
aatatctgga tagcatatgc tatcctaatc tatatctggg 2580tagcataggc tatcctaatc
tatatctggg tagcatatgc tatcctaatc tatatctggg 2640tagtatatgc tatcctaatt
tatatctggg tagcataggc tatcctaatc tatatctggg 2700tagcatatgc tatcctaatc
tatatctggg tagtatatgc tatcctaatc tgtatccggg 2760tagcatatgc tatcctaata
gagattaggg tagtatatgc tatcctaatt tatatctggg 2820tagcatatac tacccaaata
tctggatagc atatgctatc ctaatctata tctgggtagc 2880atatgctatc ctaatctata
tctgggtagc ataggctatc ctaatctata tctgggtagc 2940atatgctatc ctaatctata
tctgggtagt atatgctatc ctaatttata tctgggtagc 3000ataggctatc ctaatctata
tctgggtagc atatgctatc ctaatctata tctgggtagt 3060atatgctatc ctaatctgta
tccgggtagc atatgctatc ctcacgatga taagctgtca 3120aacatgagaa ttaattcttg
aagacgaaag ggcctcgtga tacgcctatt tttataggtt 3180aatgtcatga taataatggt
ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc 3240ggaaccccta tttgtttatt
tttctaaata cattcaaata tgtatccgct catgagacaa 3300taaccctgat aaatgcttca
ataatattga aaaaggaaga gtatgagtat tcaacatttc 3360cgtgtcgccc ttattccctt
ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 3420acgctggtga aagtaaaaga
tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 3480ctggatctca acagcggtaa
gatccttgag agttttcgcc ccgaagaacg ttttccaatg 3540atgagcactt ttaaagttct
gctatgtggc gcggtattat cccgtgttga cgccgggcaa 3600gagcaactcg gtcgccgcat
acactattct cagaatgact tggttgagta ctcaccagtc 3660acagaaaagc atcttacgga
tggcatgaca gtaagagaat tatgcagtgc tgccataacc 3720atgagtgata acactgcggc
caacttactt ctgacaacga tcggaggacc gaaggagcta 3780accgcttttt tgcacaacat
gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 3840ctgaatgaag ccataccaaa
cgacgagcgt gacaccacga tgcctgcagc aatggcaaca 3900acgttgcgca aactattaac
tggcgaacta cttactctag cttcccggca acaattaata 3960gactggatgg aggcggataa
agttgcagga ccacttctgc gctcggccct tccggctggc 4020tggtttattg ctgataaatc
tggagccggt gagcgtgggt ctcgcggtat cattgcagca 4080ctggggccag atggtaagcc
ctcccgtatc gtagttatct acacgacggg gagtcaggca 4140actatggatg aacgaaatag
acagatcgct gagataggtg cctcactgat taagcattgg 4200taactgtcag accaagttta
ctcatatata ctttagattg atttaaaact tcatttttaa 4260tttaaaagga tctaggtgaa
gatccttttt gataatctca tgaccaaaat cccttaacgt 4320gagttttcgt tccactgagc
gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 4380cctttttttc tgcgcgtaat
ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 4440gtttgtttgc cggatcaaga
gctaccaact ctttttccga aggtaactgg cttcagcaga 4500gcgcagatac caaatactgt
ccttctagtg tagccgtagt taggccacca cttcaagaac 4560tctgtagcac cgcctacata
cctcgctctg ctaatcctgt taccagtggc tgctgccagt 4620ggcgataagt cgtgtcttac
cgggttggac tcaagacgat agttaccgga taaggcgcag 4680cggtcgggct gaacgggggg
ttcgtgcaca cagcccagct tggagcgaac gacctacacc 4740gaactgagat acctacagcg
tgagcattga gaaagcgcca cgcttcccga agggagaaag 4800gcggacaggt atccggtaag
cggcagggtc ggaacaggag agcgcacgag ggagcttcca 4860gggggaaacg cctggtatct
ttatagtcct gtcgggtttc gccacctctg acttgagcgt 4920cgatttttgt gatgctcgtc
aggggggcgg agcctatgga aaaacgccag caacgcggcc 4980tttttacggt tcctggcctt
ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 5040cctgattctg tggataaccg
tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 5100cgaacgaccg agcgcagcga
gtcagtgagc gaggaagc 51385536DNAArtificial
Sequenceprimer used to generate Fc-fused KAAG1 fragment 55gtaagcaagc
ttaggccgct gggacagcgg aggtgc
365633DNAArtificial Sequenceprimer used to generate Fc-fused KAAG1
fragment 56gtaagcaagc ttggcagcag cgccaggtcc agc
335733DNAArtificial SequenceOGS1773 primer 57gtaagcagcg ctgtggctgc
accatctgtc ttc 335835DNAArtificial
SequenceOGS1774 primer 58gtaagcgcta gcctaacact ctcccctgtt gaagc
3559321DNAHomo sapiens 59gctgtggctg caccatctgt
cttcatcttc ccgccatctg atgagcagtt gaaatctgga 60actgcctctg ttgtgtgcct
gctgaataac ttctatccca gagaggccaa agtacagtgg 120aaggtggata acgccctcca
atcgggtaac tcccaggaga gtgtcacaga gcaggacagc 180aaggacagca cctacagcct
cagcagcacc ctgacgctga gcaaagcaga ctacgagaaa 240cacaaagtct acgcctgcga
agtcacccat cagggcctga gctcgcccgt cacaaagagc 300ttcaacaggg gagagtgtta g
32160106PRTHomo sapiens
60Ala Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 1
5 10 15 Leu Lys Ser Gly
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 20
25 30 Pro Arg Glu Ala Lys Val Gln Trp Lys
Val Asp Asn Ala Leu Gln Ser 35 40
45 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
Ser Thr 50 55 60
Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 65
70 75 80 His Lys Val Tyr Ala
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 85
90 95 Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
100 105 616385DNAArtificial
SequencepTTVK1 expression plasmid 61cttgagccgg cggatggtcg aggtgaggtg
tggcaggctt gagatccagc tgttggggtg 60agtactccct ctcaaaagcg ggcattactt
ctgcgctaag attgtcagtt tccaaaaacg 120aggaggattt gatattcacc tggcccgatc
tggccataca cttgagtgac aatgacatcc 180actttgcctt tctctccaca ggtgtccact
cccaggtcca agtttaaacg gatctctagc 240gaattcatga actttctgct gtcttgggtg
cattggagcc ttgccttgct gctctacctc 300caccatgcca agtggtccca ggcttgagac
ggagcttaca gcgctgtggc tgcaccatct 360gtcttcatct tcccgccatc tgatgagcag
ttgaaatctg gaactgcctc tgttgtgtgc 420ctgctgaata acttctatcc cagagaggcc
aaagtacagt ggaaggtgga taacgccctc 480caatcgggta actcccagga gagtgtcaca
gagcaggaca gcaaggacag cacctacagc 540ctcagcagca ccctgacgct gagcaaagca
gactacgaga aacacaaagt ctacgcctgc 600gaagtcaccc atcagggcct gagctcgccc
gtcacaaaga gcttcaacag gggagagtgt 660tagggtaccg cggccgcttc gaatgagatc
ccccgacctc gacctctggc taataaagga 720aatttatttt cattgcaata gtgtgttgga
attttttgtg tctctcactc ggaaggacat 780atgggagggc aaatcatttg gtcgagatcc
ctcggagatc tctagctaga gccccgccgc 840cggacgaact aaacctgact acggcatctc
tgccccttct tcgcggggca gtgcatgtaa 900tcccttcagt tggttggtac aacttgccaa
ctgggccctg ttccacatgt gacacggggg 960gggaccaaac acaaaggggt tctctgactg
tagttgacat ccttataaat ggatgtgcac 1020atttgccaac actgagtggc tttcatcctg
gagcagactt tgcagtctgt ggactgcaac 1080acaacattgc ctttatgtgt aactcttggc
tgaagctctt acaccaatgc tgggggacat 1140gtacctccca ggggcccagg aagactacgg
gaggctacac caacgtcaat cagaggggcc 1200tgtgtagcta ccgataagcg gaccctcaag
agggcattag caatagtgtt tataaggccc 1260ccttgttaac cctaaacggg tagcatatgc
ttcccgggta gtagtatata ctatccagac 1320taaccctaat tcaatagcat atgttaccca
acgggaagca tatgctatcg aattagggtt 1380agtaaaaggg tcctaaggaa cagcgatatc
tcccacccca tgagctgtca cggttttatt 1440tacatggggt caggattcca cgagggtagt
gaaccatttt agtcacaagg gcagtggctg 1500aagatcaagg agcgggcagt gaactctcct
gaatcttcgc ctgcttcttc attctccttc 1560gtttagctaa tagaataact gctgagttgt
gaacagtaag gtgtatgtga ggtgctcgaa 1620aacaaggttt caggtgacgc ccccagaata
aaatttggac ggggggttca gtggtggcat 1680tgtgctatga caccaatata accctcacaa
accccttggg caataaatac tagtgtagga 1740atgaaacatt ctgaatatct ttaacaatag
aaatccatgg ggtggggaca agccgtaaag 1800actggatgtc catctcacac gaatttatgg
ctatgggcaa cacataatcc tagtgcaata 1860tgatactggg gttattaaga tgtgtcccag
gcagggacca agacaggtga accatgttgt 1920tacactctat ttgtaacaag gggaaagaga
gtggacgccg acagcagcgg actccactgg 1980ttgtctctaa cacccccgaa aattaaacgg
ggctccacgc caatggggcc cataaacaaa 2040gacaagtggc cactcttttt tttgaaattg
tggagtgggg gcacgcgtca gcccccacac 2100gccgccctgc ggttttggac tgtaaaataa
gggtgtaata acttggctga ttgtaacccc 2160gctaaccact gcggtcaaac cacttgccca
caaaaccact aatggcaccc cggggaatac 2220ctgcataagt aggtgggcgg gccaagatag
gggcgcgatt gctgcgatct ggaggacaaa 2280ttacacacac ttgcgcctga gcgccaagca
cagggttgtt ggtcctcata ttcacgaggt 2340cgctgagagc acggtgggct aatgttgcca
tgggtagcat atactaccca aatatctgga 2400tagcatatgc tatcctaatc tatatctggg
tagcataggc tatcctaatc tatatctggg 2460tagcatatgc tatcctaatc tatatctggg
tagtatatgc tatcctaatt tatatctggg 2520tagcataggc tatcctaatc tatatctggg
tagcatatgc tatcctaatc tatatctggg 2580tagtatatgc tatcctaatc tgtatccggg
tagcatatgc tatcctaata gagattaggg 2640tagtatatgc tatcctaatt tatatctggg
tagcatatac tacccaaata tctggatagc 2700atatgctatc ctaatctata tctgggtagc
atatgctatc ctaatctata tctgggtagc 2760ataggctatc ctaatctata tctgggtagc
atatgctatc ctaatctata tctgggtagt 2820atatgctatc ctaatttata tctgggtagc
ataggctatc ctaatctata tctgggtagc 2880atatgctatc ctaatctata tctgggtagt
atatgctatc ctaatctgta tccgggtagc 2940atatgctatc ctcacgatga taagctgtca
aacatgagaa ttaattcttg aagacgaaag 3000ggcctcgtga tacgcctatt tttataggtt
aatgtcatga taataatggt ttcttagacg 3060tcaggtggca cttttcgggg aaatgtgcgc
ggaaccccta tttgtttatt tttctaaata 3120cattcaaata tgtatccgct catgagacaa
taaccctgat aaatgcttca ataatattga 3180aaaaggaaga gtatgagtat tcaacatttc
cgtgtcgccc ttattccctt ttttgcggca 3240ttttgccttc ctgtttttgc tcacccagaa
acgctggtga aagtaaaaga tgctgaagat 3300cagttgggtg cacgagtggg ttacatcgaa
ctggatctca acagcggtaa gatccttgag 3360agttttcgcc ccgaagaacg ttttccaatg
atgagcactt ttaaagttct gctatgtggc 3420gcggtattat cccgtgttga cgccgggcaa
gagcaactcg gtcgccgcat acactattct 3480cagaatgact tggttgagta ctcaccagtc
acagaaaagc atcttacgga tggcatgaca 3540gtaagagaat tatgcagtgc tgccataacc
atgagtgata acactgcggc caacttactt 3600ctgacaacga tcggaggacc gaaggagcta
accgcttttt tgcacaacat gggggatcat 3660gtaactcgcc ttgatcgttg ggaaccggag
ctgaatgaag ccataccaaa cgacgagcgt 3720gacaccacga tgcctgcagc aatggcaaca
acgttgcgca aactattaac tggcgaacta 3780cttactctag cttcccggca acaattaata
gactggatgg aggcggataa agttgcagga 3840ccacttctgc gctcggccct tccggctggc
tggtttattg ctgataaatc tggagccggt 3900gagcgtgggt ctcgcggtat cattgcagca
ctggggccag atggtaagcc ctcccgtatc 3960gtagttatct acacgacggg gagtcaggca
actatggatg aacgaaatag acagatcgct 4020gagataggtg cctcactgat taagcattgg
taactgtcag accaagttta ctcatatata 4080ctttagattg atttaaaact tcatttttaa
tttaaaagga tctaggtgaa gatccttttt 4140gataatctca tgaccaaaat cccttaacgt
gagttttcgt tccactgagc gtcagacccc 4200gtagaaaaga tcaaaggatc ttcttgagat
cctttttttc tgcgcgtaat ctgctgcttg 4260caaacaaaaa aaccaccgct accagcggtg
gtttgtttgc cggatcaaga gctaccaact 4320ctttttccga aggtaactgg cttcagcaga
gcgcagatac caaatactgt ccttctagtg 4380tagccgtagt taggccacca cttcaagaac
tctgtagcac cgcctacata cctcgctctg 4440ctaatcctgt taccagtggc tgctgccagt
ggcgataagt cgtgtcttac cgggttggac 4500tcaagacgat agttaccgga taaggcgcag
cggtcgggct gaacgggggg ttcgtgcaca 4560cagcccagct tggagcgaac gacctacacc
gaactgagat acctacagcg tgagcattga 4620gaaagcgcca cgcttcccga agggagaaag
gcggacaggt atccggtaag cggcagggtc 4680ggaacaggag agcgcacgag ggagcttcca
gggggaaacg cctggtatct ttatagtcct 4740gtcgggtttc gccacctctg acttgagcgt
cgatttttgt gatgctcgtc aggggggcgg 4800agcctatgga aaaacgccag caacgcggcc
tttttacggt tcctggcctt ttgctggcct 4860tttgctcaca tgttctttcc tgcgttatcc
cctgattctg tggataaccg tattaccgcc 4920tttgagtgag ctgataccgc tcgccgcagc
cgaacgaccg agcgcagcga gtcagtgagc 4980gaggaagcgg aagagcgccc aatacgcaaa
ccgcctctcc ccgcgcgttg gccgattcat 5040taatgcagct ggcacgacag gtttcccgac
tggaaagcgg gcagtgagcg caacgcaatt 5100aatgtgagtt agctcactca ttaggcaccc
caggctttac actttatgct tccggctcgt 5160atgttgtgtg gaattgtgag cggataacaa
tttcacacag gaaacagcta tgaccatgat 5220tacgccaagc tctagctaga ggtcgaccaa
ttctcatgtt tgacagctta tcatcgcaga 5280tccgggcaac gttgttgcat tgctgcaggc
gcagaactgg taggtatggc agatctatac 5340attgaatcaa tattggcaat tagccatatt
agtcattggt tatatagcat aaatcaatat 5400tggctattgg ccattgcata cgttgtatct
atatcataat atgtacattt atattggctc 5460atgtccaata tgaccgccat gttgacattg
attattgact agttattaat agtaatcaat 5520tacggggtca ttagttcata gcccatatat
ggagttccgc gttacataac ttacggtaaa 5580tggcccgcct ggctgaccgc ccaacgaccc
ccgcccattg acgtcaataa tgacgtatgt 5640tcccatagta acgccaatag ggactttcca
ttgacgtcaa tgggtggagt atttacggta 5700aactgcccac ttggcagtac atcaagtgta
tcatatgcca agtccgcccc ctattgacgt 5760caatgacggt aaatggcccg cctggcatta
tgcccagtac atgaccttac gggactttcc 5820tacttggcag tacatctacg tattagtcat
cgctattacc atggtgatgc ggttttggca 5880gtacaccaat gggcgtggat agcggtttga
ctcacgggga tttccaagtc tccaccccat 5940tgacgtcaat gggagtttgt tttggcacca
aaatcaacgg gactttccaa aatgtcgtaa 6000taaccccgcc ccgttgacgc aaatgggcgg
taggcgtgta cggtgggagg tctatataag 6060cagagctcgt ttagtgaacc gtcagatcct
cactctcttc cgcatcgctg tctgcgaggg 6120ccagctgttg ggctcgcggt tgaggacaaa
ctcttcgcgg tctttccagt actcttggat 6180cggaaacccg tcggcctccg aacggtactc
cgccaccgag ggacctgagc gagtccgcat 6240cgaccggatc ggaaaacctc tcgagaaagg
cgtctaacca gtcacagtcg caaggtaggc 6300tgagcaccgt ggcgggcggc agcgggtggc
ggtcggggtt gtttctggcg gaggtgctgc 6360tgatgatgta attaaagtag gcggt
63856243DNAArtificial SequencePrimer
specific for the light chain variable region of the 3D3 antibody
62atgccaagtg gtcccaggct gacattgtga tgacccagtc tcc
436343DNAArtificial SequencePrimer specific for the light chain variable
region of the 3G10 antibody 63atgccaagtg gtcccaggct gatgttttga
tgacccaaac tcc 436443DNAArtificial SequencePrimer
specific for the light chain variable region of the 3C4 antibody
64atgccaagtg gtcccaggct gacatcgtta tgtctcagtc tcc
436532DNAArtificial Sequencereverse primer to amplify the 3D3, 3G10 and
3C4 antibody light chains 65gggaagatga agacagatgg tgcagccaca gc
326650DNAArtificial SequenceOGS1769 primer
66gtaagcgcta gcgcctcaac gaagggccca tctgtctttc ccctggcccc
506737DNAArtificial SequenceOGS1770 primer 67gtaagcgaat tcacaagatt
tgggctcaac tttcttg 3768309DNAHomo sapiens
68gcctccacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg
60ggcacagcag ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg
120tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca
180ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc
240tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc
300aaatcttgt
30969103PRTHomo sapiens 69Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
Pro Ser Ser Lys 1 5 10
15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30 Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35
40 45 Gly Val His Thr Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser 50 55
60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
Thr Gln Thr 65 70 75
80 Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95 Lys Val Glu Pro
Lys Ser Cys 100 705379DNAArtificial SequencepYD15
plasmid 70cttgagccgg cggatggtcg aggtgaggtg tggcaggctt gagatccagc
tgttggggtg 60agtactccct ctcaaaagcg ggcattactt ctgcgctaag attgtcagtt
tccaaaaacg 120aggaggattt gatattcacc tggcccgatc tggccataca cttgagtgac
aatgacatcc 180actttgcctt tctctccaca ggtgtccact cccaggtcca agtttgccgc
caccatggag 240acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac
tggcggagac 300ggagcttacg ggcccatctg tctttcccct ggccccctcc tccaagagca
cctctggggg 360cacagcggcc ctgggctgcc tggtcaagga ctacttcccc gaaccggtga
cggtgtcgtg 420gaactcaggc gccctgacca gcggcgtgca caccttcccg gctgtcctac
agtcctcagg 480actctactcc ctcagcagcg tggtgaccgt gccctccagc agcttgggca
cccagaccta 540catctgcaac gtgaatcaca agcccagcaa caccaaggtg gacaagaaag
ttgagcccaa 600atcttgtgaa ttcactcaca catgcccacc gtgcccagca cctgaactcc
tggggggacc 660gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc
ggacccctga 720ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt
tcaactggta 780cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc
agtacaacag 840cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga
atggcaagga 900gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa
ccatctccaa 960agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc
gggatgagct 1020gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca
gcgacatcgc 1080cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc
ctcccgtgct 1140ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga
gcaggtggca 1200gcaggggaac gtcttctcat gctccgtgat gcatgaggct ctgcacaacc
actacacgca 1260gaagagcctc tccctgtctc ccgggaaatg atcccccgac ctcgacctct
ggctaataaa 1320ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca
ctcggaagga 1380catatgggag ggcaaatcat ttggtcgaga tccctcggag atctctagct
agagccccgc 1440cgccggacga actaaacctg actacggcat ctctgcccct tcttcgcggg
gcagtgcatg 1500taatcccttc agttggttgg tacaacttgc caactgaacc ctaaacgggt
agcatatgct 1560tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata
tgttacccaa 1620cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac
agcgatgtag 1680gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt
acacacactt 1740gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg
ctgagagcac 1800ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata
gcatatgcta 1860tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
gcatatgcta 1920tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
gcataggcta 1980tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
gtatatgcta 2040tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta
gtatatgcta 2100tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat
atgctatcct 2160aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat
aggctatcct 2220aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
atgctatcct 2280aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
atgctatcct 2340aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
atgctatcct 2400cacgatgata agctgtcaaa catgagaatt aattcttgaa gacgaaaggg
cctcgtgata 2460cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
aggtggcact 2520tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
ttcaaatatg 2580tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
aaggaagagt 2640atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
ttgccttcct 2700gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
gttgggtgca 2760cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
ttttcgcccc 2820gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
ggtattatcc 2880cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
gaatgacttg 2940gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
aagagaatta 3000tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
gacaacgatc 3060ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
aactcgcctt 3120gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
caccacgatg 3180cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
tactctagct 3240tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
acttctgcgc 3300tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
gcgtgggtct 3360cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
agttatctac 3420acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
gataggtgcc 3480tcactgatta agcattggta actgtcagac caagtttact catatatact
ttagattgat 3540ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
taatctcatg 3600accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
agaaaagatc 3660aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
aacaaaaaaa 3720ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
ttttccgaag 3780gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
gccgtagtta 3840ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
aatcctgtta 3900ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
aagacgatag 3960ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
gcccagcttg 4020gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
aagcgccacg 4080cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
aacaggagag 4140cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
cgggtttcgc 4200cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
cctatggaaa 4260aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
tgctcacatg 4320ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
tgagtgagct 4380gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
ggaagcgtac 4440atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt
gactagttat 4500taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
ccgcgttaca 4560taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
attgacgtca 4620ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
tcaatgggtg 4680gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
gccaagtccg 4740ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
gtacatgacc 4800ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
taccatggtg 4860atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
gggatttcca 4920agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
acgggacttt 4980ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
tgtacggtgg 5040gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcctcactct
cttccgcatc 5100gctgtctgcg agggccagct gttgggctcg cggttgagga caaactcttc
gcggtctttc 5160cagtactctt ggatcggaaa cccgtcggcc tccgaacggt actccgccac
cgagggacct 5220gagcgagtcc gcatcgaccg gatcggaaaa cctctcgaga aaggcgtcta
accagtcaca 5280gtcgcaaggt aggctgagca ccgtggcggg cggcagcggg tggcggtcgg
ggttgtttct 5340ggcggaggtg ctgctgatga tgtaattaaa gtaggcggt
53797143DNAArtificial SequencePrimer specific for the heavy
chain variable region of the 3D3 and 3G10 antibodies 71gggttccagg
ttccactggc gaggttcagc tgcagcagtc tgt
437243DNAArtificial SequencePrimer specific for the heavy chain variable
region of the 3C4 antibody 72gggttccagg ttccactggc gaggtgcagc
ttcaggagtc agg 437338DNAArtificial Sequencereverse
primer to amplify the 3D3, 3G10 and 3C4 antibody heavy chains
73ggggccaggg gaaagacaga tgggcccttc gttgaggc
387417PRTArtificial Sequencelight chain CDR1 consensus version
1MISC_FEATURE(1)..(1)Xaa is a basic amino acidMISC_FEATURE(4)..(4)Xaa is
a basic amino acidMISC_FEATURE(8)..(8)Xaa is His, Tyr or
AsnMISC_FEATURE(9)..(9)Xaa is Ser, Thr, Asn or
ArgMISC_FEATURE(10)..(10)Xaa is absent, Ser or
AsnMISC_FEATURE(11)..(11)Xaa is Asp, Phe or AsnMISC_FEATURE(12)..(12)Xaa
is Gly or GlnMISC_FEATURE(13)..(13)Xaa is Lys, Leu or
AsnMISC_FEATURE(14)..(14)Xaa is Thr or AsnMISC_FEATURE(15)..(15)Xaa is an
aromatic amino acidMISC_FEATURE(17)..(17)Xaa is Ala, Asn, Glu or Tyr
74Xaa Ser Ser Xaa Ser Leu Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Leu 1
5 10 15 Xaa
7511PRTArtificial Sequencelight chain CDR1 consensus version
2MISC_FEATURE(6)..(6)Xaa is an hydrophobic amino
acidMISC_FEATURE(7)..(7)Xaa is Gly or HisMISC_FEATURE(8)..(8)Xaa is Thr,
Asn or ArgMISC_FEATURE(9)..(9)Xaa is Phe, Tyr or
AlaMISC_FEATURE(10)..(10)Xaa is an hydrophobic amino
acidMISC_FEATURE(11)..(11)Xaa is Asn or Ala 75Lys Ala Ser Gln Asp Xaa Xaa
Xaa Xaa Xaa Xaa 1 5 10
767PRTArtificial Sequencelight chain CDR2 consensus version
1MISC_FEATURE(2)..(2)Xaa is Ala or GlyMISC_FEATURE(5)..(5)Xaa is Arg or
ThrMISC_FEATURE(6)..(6)Xaa is Glu, Lys or Ala 76Phe Xaa Ser Thr Xaa Xaa
Ser 1 5 777PRTArtificial Sequencelight chain CDR2
consensus version 2MISC_FEATURE(1)..(1)Xaa is Leu or
LysMISC_FEATURE(4)..(4)Xaa is a basic amino acidMISC_FEATURE(5)..(5)Xaa
is Leu or ArgMISC_FEATURE(6)..(6)Xaa is Asp or Phe 77Xaa Val Ser Xaa Xaa
Xaa Ser 1 5 787PRTArtificial Sequencelight chain
CDR2 consensus version 3MISC_FEATURE(1)..(1)Xaa is a basic amino
acidMISC_FEATURE(7)..(7)Xaa is Asp or Ala 78Xaa Ala Asn Arg Leu Val Xaa 1
5 799PRTArtificial Sequencelight chain CDR3
consensus version 1MISC_FEATURE(1)..(1)Xaa is Gln or
LeuMISC_FEATURE(3)..(3)Xaa is an aromatic amino
acidMISC_FEATURE(4)..(4)Xaa is Asp, Phe or TyrMISC_FEATURE(5)..(5)Xaa is
Glu, Ala, Asn or SerMISC_FEATURE(6)..(6)Xaa is Ile, Phe or Thr 79Xaa Gln
Xaa Xaa Xaa Xaa Pro Leu Thr 1 5
809PRTArtificial Sequencelight chain CDR3 consensus version
2MISC_FEATURE(4)..(4)Xaa is an aromatic amino acidMISC_FEATURE(5)..(5)Xaa
is Asn or SerMISC_FEATURE(6)..(6)Xaa is Ile or Thr 80Gln Gln His Xaa Xaa
Xaa Pro Leu Thr 1 5 819PRTArtificial
Sequencelight chain CDR3 consensus version 3MISC_FEATURE(1)..(1)Xaa is an
aromatic amino acidMISC_FEATURE(4)..(4)Xaa is a neutral hydrophilic amino
acidMISC_FEATURE(6)..(6)Xaa is Phe or ValMISC_FEATURE(8)..(8)Xaa is Arg
or Leu 81Xaa Gln Gly Xaa His Xaa Pro Xaa Thr 1 5
8210PRTArtificial Sequenceheavy chain CDR1
consensusMISC_FEATURE(3)..(3)Xaa is Thr, Ile or
LysMISC_FEATURE(5)..(5)Xaa is a neutral hydrophilic amino
acidMISC_FEATURE(6)..(6)Xaa is an acidic amino
acidMISC_FEATURE(8)..(8)Xaa is Glu, Asn or AspMISC_FEATURE(9)..(9)Xaa is
an hydrophobic amino acid 82Gly Tyr Xaa Phe Xaa Xaa Tyr Xaa Xaa His 1
5 10 8310PRTArtificial Sequenceheavy chain
CDR2 consensus version 1MISC_FEATURE(1)..(1)Xaa is Val or
GlyMISC_FEATURE(2)..(2)Xaa is an hydrophobic amino
acidMISC_FEATURE(5)..(5)Xaa is Ala, Gly or GluMISC_FEATURE(8)..(8)Xaa is
Arg, Gly, Asp, Ala, Ser, Asn or ValMISC_FEATURE(10)..(10)Xaa is an
hydrophobic amino acid 83Xaa Xaa Asp Pro Xaa Thr Gly Xaa Thr Xaa 1
5 10 8410PRTArtificial Sequenceheavy chain CDR2
consensus version 2MISC_FEATURE(2)..(2)Xaa is an hydrophobic amino
acidMISC_FEATURE(5)..(5)Xaa is Ala, Glu or GlyMISC_FEATURE(8)..(8)Xaa is
Arg, Gly, Ala, Ser, Asn, Val or Asp 84Val Xaa Asp Pro Xaa Thr Gly Xaa Thr
Ala 1 5 10 859PRTArtificial
Sequenceheavy chain CDR2 consensus version 3MISC_FEATURE(3)..(3)Xaa is
Ser or AsnMISC_FEATURE(4)..(4)Xaa is an aromatic amino
acidMISC_FEATURE(5)..(5)Xaa is Asp, Glu or AsnMISC_FEATURE(7)..(7)Xaa is
Asp or HisMISC_FEATURE(8)..(8)Xaa is Tyr, Ser or
AsnMISC_FEATURE(9)..(9)Xaa is Asp, Glu or Asn 85Tyr Ile Xaa Xaa Xaa Gly
Xaa Xaa Xaa 1 5 8610PRTArtificial
Sequenceheavy chain CDR2 consensus version 4MISC_FEATURE(1)..(1)Xaa is
Asn or TyrMISC_FEATURE(7)..(7)Xaa is Glu, Asp or Asn 86Xaa Ile Asn Pro
Tyr Asn Xaa Val Thr Glu 1 5 10
8710PRTArtificial Sequenceheavy chain CDR2 consensus version
5MISC_FEATURE(5)..(5)Xaa is Asn or TyrMISC_FEATURE(8)..(8)Xaa is Gly or
ThrMISC_FEATURE(9)..(9)Xaa is Ile or Thr 87Asp Ile Asn Pro Xaa Tyr Gly
Xaa Xaa Thr 1 5 10 886PRTArtificial
Sequenceheavy chain CDR3 consensus version 1MISC_FEATURE(2)..(2)Xaa is
Gly or SerMISC_FEATURE(3)..(3)Xaa is Tyr or HisMISC_FEATURE(4)..(4)Xaa is
Ala or Ser 88Met Xaa Xaa Xaa Asp Tyr 1 5
897PRTArtificial Sequenceheavy chain CDR3 consensus version
2MISC_FEATURE(2)..(2)Xaa is Gly or SerMISC_FEATURE(5)..(5)Xaa is absent
or Met 89Ile Xaa Tyr Ala Xaa Asp Tyr 1 5
907PRTArtificial Sequenceheavy chain CDR3 consensus version
3MISC_FEATURE(2)..(2)Xaa is Arg or TrpMISC_FEATURE(3)..(3)Xaa is an
aromatic amino acidMISC_FEATURE(7)..(7)Xaa is a basic amino acid 90Ala
Xaa Xaa Gly Leu Arg Xaa 1 5 9117PRTArtificial
SequenceExemplary embodiment of a light chain CDR1MISC_FEATURE(8)..(8)Xaa
is Asn or HisMISC_FEATURE(9)..(9)Xaa is Ser or
ThrMISC_FEATURE(10)..(10)Xaa is Ser, Asn or AspMISC_FEATURE(11)..(11)Xaa
is Asn or GlyMISC_FEATURE(12)..(12)Xaa is Gln, Asn or
LysMISC_FEATURE(13)..(13)Xaa is Lys or Leu 91Lys Ser Ser Gln Ser Leu Leu
Xaa Xaa Xaa Xaa Xaa Xaa Asn Tyr Leu 1 5
10 15 Ala 9211PRTArtificial SequenceExemplary
embodiment of a light chain CDR1MISC_FEATURE(8)..(8)Xaa is Asn or
ThrMISC_FEATURE(9)..(9)Xaa is Tyr or Phe 92Lys Ala Ser Gln Asp Ile His
Xaa Xaa Leu Asn 1 5 10
937PRTArtificial SequenceExemplary embodiment of a light chain CDR2 93Phe
Ala Ser Thr Arg Glu Ser 1 5 947PRTArtificial
SequenceExemplary embodiment of a light chain CDR2 94Leu Val Ser Lys Leu
Asp Ser 1 5 957PRTArtificial SequenceExemplary
embodiment of a light chain CDR2 95Arg Ala Asn Arg Leu Val Asp 1
5 969PRTArtificial SequenceExemplary embodiment of a light
chain CDR3 96Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
979PRTArtificial SequenceExemplary embodiment of a light chain
CDR3MISC_FEATURE(1)..(1)Xaa is Trp or LeuMISC_FEATURE(3)..(3)Xaa is Tyr
or GlyMISC_FEATURE(4)..(4)Xaa is Asp or ThrMISC_FEATURE(5)..(5)Xaa is
Ala, Glu or His 97Xaa Gln Xaa Xaa Xaa Phe Pro Arg Thr 1 5
9810PRTArtificial SequenceExemplary embodiment of a heavy
chain CDR1MISC_FEATURE(3)..(3)Xaa is The or IleMISC_FEATURE(6)..(6)Xaa is
Asp or GluMISC_FEATURE(8)..(8)Xaa is Glu or AsnMISC_FEATURE(9)..(9)Xaa is
Met, Ile or Val 98Gly Tyr Xaa Phe Thr Xaa Tyr Xaa Xaa His 1
5 10 9911PRTArtificial SequenceExemplary embodiment of
a heavy chain CDR1MISC_FEATURE(3)..(3)Xaa is Thr or Ser 99Gly Phe Xaa Ile
Thr Ser Gly Tyr Gly Trp His 1 5 10
10010PRTArtificial SequenceExemplary embodiment of a heavy chain
CDR2MISC_FEATURE(1)..(1)Xaa is Val, Asn or GlyMISC_FEATURE(2)..(2)Xaa is
Ile or LeuMISC_FEATURE(5)..(5)Xaa is Glu, Ala or
GlyMISC_FEATURE(6)..(6)Xaa is Thr or Tyrmisc_feature(8)..(8)Xaa can be
any naturally occurring amino acid 100Xaa Xaa Asp Pro Xaa Xaa Gly Xaa Thr
Ala 1 5 10 1016PRTArtificial
SequenceExemplary embodiment of a heavy chain CDR2MISC_FEATURE(3)..(3)Xaa
is Asn or SerMISC_FEATURE(4)..(4)Xaa is Phe or TyrMISC_FEATURE(5)..(5)Xaa
is Asn or Asp 101Tyr Ile Xaa Xaa Xaa Gly 1 5
1026PRTArtificial SequenceExemplary embodiment of a heavy chain
CDR3MISC_FEATURE(4)..(4)Xaa is Ser or Ala 102Met Gly Tyr Xaa Asp Tyr 1
5 10310PRTArtificial SequenceExemplary embodiment of a
heavy chain CDR3 103Ala Ser Ser Tyr Asp Gly Phe Leu Ala Tyr 1
5 10 1047PRTArtificial SequenceExemplary embodiment
of a heavy chain CDR3MISC_FEATURE(2)..(2)Xaa is Arg or
TrpMISC_FEATURE(3)..(3)Xaa is Trp or PheMISC_FEATURE(7)..(7)Xaa is Gln or
Asn 104Ala Xaa Xaa Gly Leu Arg Xaa 1 5
105112PRTArtificial SequenceAmino acid sequence of the 3z1A02 light chain
105Asp Ala Val Met Thr Gln Ile Pro Leu Thr Leu Ser Val Thr Ile Gly 1
5 10 15 Gln Pro Ala Ser
Leu Ser Cys Lys Ser Ser Gln Ser Leu Leu His Ser 20
25 30 Asp Gly Lys Thr Tyr Leu Asn Trp Leu
Leu Gln Arg Pro Gly Gln Ser 35 40
45 Pro Lys Arg Leu Ile Ser Leu Val Ser Lys Leu Asp Ser Gly
Val Pro 50 55 60
Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65
70 75 80 Ser Arg Val Glu Ala
Glu Asp Leu Gly Leu Tyr Tyr Cys Trp Gln Gly 85
90 95 Thr His Phe Pro Arg Thr Phe Ala Gly Gly
Thr Asn Leu Glu Ile Lys 100 105
110 106112PRTArtificial SequenceAmino acid sequence of the
3z1F06 light chain 106Ser Ile Val Met Thr Gln Thr Pro Leu Thr Leu Ser Val
Thr Ile Gly 1 5 10 15
Gln Pro Ala Ser Ile Thr Cys Lys Ser Ser Gln Ser Leu Leu Tyr Ser
20 25 30 Asp Gly Lys Thr
Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln Ser 35
40 45 Pro Lys Arg Leu Ile Ser Leu Val Ser
Lys Leu Asp Ser Gly Val Pro 50 55
60 Asp Gly Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr
Leu Lys Ile 65 70 75
80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr Tyr Cys Trp Gln Gly
85 90 95 Thr His Phe Pro
Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100
105 110 107112PRTArtificial SequenceAmino
acid sequence of the 3z1E08 light chain 107Asp Ala Val Met Thr Gln Ile
Pro Leu Thr Leu Ser Val Thr Ile Gly 1 5
10 15 Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln
Ser Leu Leu His Ser 20 25
30 Asp Gly Lys Thr Tyr Leu Asn Trp Leu Leu Gln Arg Pro Gly Gln
Ser 35 40 45 Pro
Lys Arg Leu Ile Tyr Leu Val Ser Lys Leu Asp Ser Gly Val Pro 50
55 60 Asp Arg Phe Thr Gly Ser
Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65 70
75 80 Ser Arg Val Glu Ala Glu Asp Leu Gly Val Tyr
Tyr Cys Trp Gln Gly 85 90
95 Thr His Phe Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105 110
108112PRTArtificial SequenceAmino acid sequence of the 3z1G10 light chain
108Asp Val Leu Met Thr Gln Thr Pro Arg Ser Leu Ser Val Ser Leu Gly 1
5 10 15 Asp Gln Ala Ser
Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 20
25 30 Asn Gly Asn Thr Tyr Leu Glu Trp Tyr
Leu Gln Lys Pro Gly Gln Pro 35 40
45 Pro Lys Val Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly
Val Pro 50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 65
70 75 80 Ser Gly Val Glu Ala
Glu Asp Leu Gly Val Tyr Tyr Cys Phe Gln Gly 85
90 95 Ser His Val Pro Leu Thr Phe Gly Ala Gly
Thr Lys Leu Glu Leu Lys 100 105
110 109112PRTArtificial SequenceAmino acid sequence of the
3z1E10 light chain 109Asp Ile Val Met Thr Gln Ala Ala Pro Ser Val Pro Val
Thr Pro Gly 1 5 10 15
Glu Ser Val Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu His Ser
20 25 30 Asn Gly Asn Thr
Tyr Leu Tyr Trp Phe Leu Gln Arg Pro Gly Gln Ser 35
40 45 Pro Gln Leu Leu Ile Tyr Arg Met Ser
Asn Leu Ala Ser Gly Val Pro 50 55
60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Ala Phe Thr
Leu Arg Ile 65 70 75
80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln His
85 90 95 Leu Glu Tyr Pro
Tyr Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100
105 110 110113PRTArtificial SequenceAmino
acid sequence of the 3z1A09 light chain 110Asp Ile Val Met Thr Gln Ser
Pro Ser Ser Leu Ala Met Ser Leu Gly 1 5
10 15 Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln
Ser Leu Leu Asn Ser 20 25
30 Asn Asn Gln Leu Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly
Gln 35 40 45 Ser
Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Lys Ser Gly Val 50
55 60 Pro Asp Arg Phe Ile Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Thr Ser Val Gln Ala Glu Asp Leu Ala Asp
Tyr Phe Cys Gln Gln 85 90
95 His Phe Asn Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
100 105 110 Lys
111113PRTArtificial SequenceAmino acid sequence of the 3z1B01 light chain
111Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Ile Ser Val Gly 1
5 10 15 Gln Lys Val Thr
Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20
25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln 35 40
45 Ser Pro Lys Leu Leu Val Phe Phe Ala Ser Thr Arg Glu Ser
Gly Val 50 55 60
Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65
70 75 80 Ile Ser Ser Val Gln
Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85
90 95 His Tyr Ser Ile Pro Leu Thr Phe Gly Ala
Gly Thr Lys Leu Glu Leu 100 105
110 Lys 112113PRTArtificial SequenceAmino acid sequence of the
3z1G05 light chain 112Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met
Ser Val Gly 1 5 10 15
Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30 Ser Asn Gln Lys
Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35
40 45 Ser Pro Lys Leu Leu Val Phe Phe Ala
Ser Thr Arg Glu Ser Gly Val 50 55
60 Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr 65 70 75
80 Ile Thr Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln
85 90 95 His Tyr Ser Ile
Pro Leu Thr Phe Gly Ser Gly Thr Lys Leu Glu Leu 100
105 110 Lys 113113PRTArtificial
SequenceAmino acid sequence of the 3z1B02 light chain 113Asp Ile Val Met
Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1 5
10 15 Gln Lys Val Thr Met Ser Cys Lys Ser
Ser Gln Ser Leu Leu Asn Ser 20 25
30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45
Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser Gly Val 50
55 60 Pro Asp Arg Phe Ile
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
Asp Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Ile Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu
Leu 100 105 110 Lys
114113PRTArtificial SequenceAmino acid sequence of the 3z1B08 light chain
114Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1
5 10 15 Gln Lys Val Thr
Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20
25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln 35 40
45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser
Gly Val 50 55 60
Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65
70 75 80 Ile Ser Ser Val Gln
Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85
90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala
Gly Thr Lys Leu Glu Leu 100 105
110 Lys 115113PRTArtificial SequenceAmino acid sequence of the
3z1G08 light chain 115Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met
Ser Val Gly 1 5 10 15
Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30 Ser Asn Gln Lys
Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35
40 45 Ser Pro Lys Leu Leu Val Tyr Phe Ala
Ser Thr Arg Glu Ser Gly Val 50 55
60 Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr 65 70 75
80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln
85 90 95 His Tyr Ser Thr
Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100
105 110 Lys 116113PRTArtificial
SequenceAmino acid sequence of the 3z1F07 light chain 116Asp Ile Val Met
Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1 5
10 15 Gln Lys Val Thr Met Ser Cys Lys Ser
Ser Gln Ser Leu Leu Asn Ser 20 25
30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45
Ser Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Glu Ser Gly Val 50
55 60 Pro Asp Arg Phe Ile
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
Asp Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu
Leu 100 105 110 Lys
117113PRTArtificial SequenceAmino acid sequence of the 3z1E09 light chain
117Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1
5 10 15 Gln Lys Val Thr
Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20
25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln 35 40
45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser
Gly Val 50 55 60
Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 65
70 75 80 Ile Thr Ser Val Gln
Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85
90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala
Gly Thr Lys Leu Glu Leu 100 105
110 Lys 118113PRTArtificial SequenceAmino acid sequence of the
3z1C03 light chain 118Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met
Ser Val Gly 1 5 10 15
Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30 Ser Asn Gln Lys
Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35
40 45 Ser Pro Lys Leu Leu Val Tyr Phe Gly
Ser Thr Arg Glu Ser Gly Val 50 55
60 Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr 65 70 75
80 Ile Ser Gly Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln
85 90 95 His Tyr Ser Thr
Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100
105 110 Lys 119113PRTArtificial
SequenceAmino acid sequence of the 3z1E12 light chain 119Asp Ile Val Met
Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1 5
10 15 Gln Lys Val Thr Met Asn Cys Lys Ser
Ser Gln Ser Leu Leu Asn Arg 20 25
30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45
Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser Gly Val 50
55 60 Pro Asp Arg Phe Ile
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
Asp Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Ile Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu
Leu 100 105 110 Lys
120113PRTArtificial SequenceAmino acid sequence of the 4z1A02 light chain
120Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1
5 10 15 Gln Lys Val Thr
Met Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Asn 20
25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln 35 40
45 Ser Pro Lys Leu Leu Leu Tyr Phe Ala Ser Thr Arg Glu Ser
Gly Val 50 55 60
Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Tyr Phe Thr Leu Thr 65
70 75 80 Ile Ser Ser Val Gln
Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85
90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala
Gly Thr Lys Leu Asp Leu 100 105
110 Lys 121113PRTArtificial SequenceAmino acid sequence of the
3z1F10 light chain 121Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Met
Ser Val Gly 1 5 10 15
Gln Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Thr
20 25 30 Ser Asn Gln Leu
Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35
40 45 Ser Pro Lys Leu Leu Val Tyr Phe Ala
Ser Thr Thr Glu Ser Gly Val 50 55
60 Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr 65 70 75
80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln
85 90 95 His Tyr Ser Thr
Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100
105 110 Lys 122113PRTArtificial
SequenceAmino acid sequence of the 3z1F04 light chain 122Asp Ile Val Met
Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 1 5
10 15 Glu Lys Val Thr Met Ser Cys Lys Ser
Ser Gln Ser Leu Leu Asn Thr 20 25
30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45
Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Ala Ser Gly Val 50
55 60 Pro Asp Arg Phe Ile
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala
Asp Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu
Leu 100 105 110 Lys
123113PRTArtificial SequenceAmino acid sequence of the 3z1B11 light chain
123Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1
5 10 15 Gln Lys Val Thr
Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20
25 30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln 35 40
45 Ser Pro Lys Leu Leu Val Tyr Phe Ala Ser Thr Arg Glu Ser
Gly Val 50 55 60
Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65
70 75 80 Ile Ser Ser Val Gln
Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln 85
90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala
Gly Thr Lys Leu Glu Leu 100 105
110 Lys 124113PRTArtificial SequenceAmino acid sequence of the
3z1D03 ight chain 124Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ala Val
Ser Ile Gly 1 5 10 15
Gln Lys Val Thr Met Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30 Asn Phe Gln Lys
Asn Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35
40 45 Ser Pro Lys Leu Leu Ile Tyr Phe Ala
Ser Thr Arg Glu Ser Ser Ile 50 55
60 Pro Asp Arg Phe Ile Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr 65 70 75
80 Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Asp Tyr Phe Cys Gln Gln
85 90 95 His Tyr Ser Thr
Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 100
105 110 Lys 125113PRTArtificial
SequenceAmino acid sequence of the 3z1C03 light chain 125Asp Ile Val Met
Thr Gln Ser Pro Ser Ser Leu Ala Met Ser Val Gly 1 5
10 15 Gln Lys Val Thr Met Ser Cys Lys Ser
Ser Gln Ser Leu Leu Asn Ser 20 25
30 Ser Asn Gln Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45
Ser Pro Lys Leu Leu Val Tyr Phe Gly Ser Thr Arg Glu Ser Gly Val 50
55 60 Pro Asp Arg Phe Ile
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65 70
75 80 Ile Ser Gly Val Gln Ala Glu Asp Leu Ala
Asp Tyr Phe Cys Gln Gln 85 90
95 His Tyr Ser Thr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu
Leu 100 105 110 Lys
126107PRTArtificial SequenceAmino acid sequence of the 3z1G12 light chain
126Asp Ile Val Met Thr Gln Ser Pro Lys Phe Met Ser Thr Ser Val Gly 1
5 10 15 Asp Arg Val Ser
Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala 20
25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly
Gln Ser Pro Glu Leu Leu Ile 35 40
45 Tyr Trp Thr Ser Thr Arg His Thr Gly Val Pro Asp Arg Phe
Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Val Gln Ala 65
70 75 80 Glu Asp Leu Ala Asp
Tyr Phe Cys Gln Gln His Tyr Ser Ile Pro Leu 85
90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
Arg 100 105 127107PRTArtificial
SequenceAmino acid sequence of the 3z1C04 light chain 127Asp Ile Val Met
Ser Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1 5
10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala
Ser Gln Asp Ile His Asn Phe 20 25
30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr
Leu Ile 35 40 45
Phe Arg Ala Asn Arg Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50
55 60 Ser Gly Ser Gly Gln
Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Phe 65 70
75 80 Glu Asp Leu Gly Ile Tyr Ser Cys Leu Gln
Tyr Asp Glu Ile Pro Leu 85 90
95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Arg 100
105 128107PRTArtificial SequenceAmino acid
sequence of the 3z1D01 light chain 128Asp Ile Lys Met Thr Gln Ser Pro Ser
Ser Met Tyr Ala Ser Leu Gly 1 5 10
15 Glu Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile His
Thr Tyr 20 25 30
Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Glu Thr Leu Ile
35 40 45 Tyr Arg Ala Asn
Arg Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50
55 60 Ser Gly Ser Gly Gln Asp Tyr Ser
Leu Thr Ile Ser Ser Leu Glu Tyr 65 70
75 80 Glu Asp Met Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp
Glu Phe Pro Leu 85 90
95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100
105 129107PRTArtificial SequenceAmino acid sequence of
the 3z1C02 light chain 129Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Tyr
Ala Ser Leu Gly 1 5 10
15 Glu Arg Val Thr Leu Thr Cys Lys Ala Ser Gln Asp Ile His Asn Tyr
20 25 30 Leu Asn Trp
Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr Leu Ile 35
40 45 His Arg Ala Asn Arg Leu Val Ala
Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser
Leu Glu Tyr 65 70 75
80 Glu Asp Leu Gly Ile Tyr Tyr Cys Leu Gln Tyr Asp Ala Phe Pro Leu
85 90 95 Thr Phe Gly Ala
Gly Thr Lys Leu Glu Leu Lys 100 105
130107PRTArtificial SequenceAmino acid sequence of the 3z1E06 light chain
130Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1
5 10 15 Glu Arg Val Thr
Leu Thr Cys Lys Ala Ser Gln Asp Ile His Asn Tyr 20
25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly
Lys Ser Pro Lys Thr Leu Ile 35 40
45 His Arg Ala Asn Arg Leu Val Ala Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60
Ser Gly Ser Gly Gln Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Tyr 65
70 75 80 Glu Asp Leu Gly Ile
Tyr Tyr Cys Leu Gln Tyr Asp Ala Phe Pro Leu 85
90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
Lys 100 105 131107PRTArtificial
SequenceAmino acid sequence of the 3z1H03 light chain 131Asp Ile Val Met
Ser Gln Ser Pro Ser Ser Met Tyr Ala Ser Leu Gly 1 5
10 15 Glu Arg Val Thr Ile Thr Cys Lys Ala
Ser Gln Asp Ile His Arg Phe 20 25
30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys Ser Pro Lys Thr
Leu Ile 35 40 45
Phe His Ala Asn Arg Leu Val Asp Gly Val Pro Ser Arg Phe Ser Gly 50
55 60 Ser Gly Ser Gly Leu
Asp Tyr Ser Leu Thr Ile Ser Ser Leu Glu Tyr 65 70
75 80 Glu Asp Met Gly Ile Tyr Phe Cys Leu Gln
Tyr Asp Ala Phe Pro Leu 85 90
95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys 100
105 132115PRTArtificial SequenceAmino acid
sequence of the 3z1A02 heavy chain 132His Glu Ile Gln Leu Gln Gln Ser Gly
Pro Glu Leu Val Lys Pro Gly 1 5 10
15 Ala Ser Val Lys Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe
Thr Asp 20 25 30
Tyr Asn Met His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp
35 40 45 Ile Gly Tyr Ile
Asn Pro Tyr Asn Asp Val Thr Glu Tyr Asn Glu Lys 50
55 60 Phe Lys Gly Arg Ala Thr Leu Thr
Ser Asp Lys Ser Ser Ser Thr Ala 65 70
75 80 Tyr Met Asp Leu Ser Ser Leu Thr Ser Asp Asp Ser
Ala Val Tyr Phe 85 90
95 Cys Ala Trp Phe Gly Leu Arg Gln Trp Gly Gln Gly Thr Leu Val Thr
100 105 110 Val Ser Thr
115 133115PRTArtificial SequenceAmino acid sequence of the 3z1F06
heavy chain 133His Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys
Pro Gly 1 5 10 15
Ala Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Ile Phe Thr Glu
20 25 30 Tyr Asn Ile His Trp
Val Lys Gln Lys Pro Gly Gln Gly Pro Glu Trp 35
40 45 Ile Gly Asn Ile Asn Pro Tyr Asn Asp
Val Thr Glu Tyr Asn Glu Lys 50 55
60 Phe Lys Gly Lys Ala Thr Leu Thr Ser Asp Lys Ala Ser
Ser Thr Ala 65 70 75
80 Tyr Met Asp Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr
85 90 95 Cys Ala Arg Trp
Gly Leu Arg Asn Trp Gly Gln Gly Thr Leu Val Thr 100
105 110 Val Ser Ala 115
134115PRTArtificial SequenceAmino acid sequence of the 3z1E08 heavy chain
134His Glu Val Gln Leu Gln Gln Ser Val Pro Glu Leu Val Lys Pro Gly 1
5 10 15 Ala Ser Val Lys
Met Ser Cys Lys Thr Ser Gly Tyr Thr Phe Thr Glu 20
25 30 Tyr Asn Met His Trp Val Lys Gln Lys
Pro Gly Gln Gly Pro Glu Trp 35 40
45 Ile Gly Asn Ile Asn Pro Tyr Asn Asn Val Thr Glu Tyr Asn
Glu Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ser Asp Lys Ser Ser Ser Thr Ala 65
70 75 80 Tyr Leu Asp Leu Ser
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Ala Arg Trp Gly Leu Arg Asn Trp Gly
Gln Gly Thr Leu Val Thr 100 105
110 Val Ser Ala 115 135114PRTArtificial SequenceAmino
acid sequence of the 3z1A09 heavy chain 135His Gln Val Gln Val Gln Gln
Pro Gly Ala Glu Leu Val Arg Pro Gly 1 5
10 15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly
Tyr Ile Phe Thr Asp 20 25
30 Tyr Glu Val His Trp Val Arg Gln Arg Pro Val His Gly Leu Glu
Trp 35 40 45 Ile
Gly Val Ile Asp Pro Glu Thr Gly Asp Thr Ala Tyr Asn Gln Lys 50
55 60 Phe Lys Gly Lys Ala Thr
Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70
75 80 Tyr Met Glu Leu Ser Ser Leu Thr Ala Glu Asp
Ser Ala Val Tyr Tyr 85 90
95 Cys Ile Gly Tyr Ala Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val
100 105 110 Ser Ser
136114PRTArtificial SequenceAmino acid sequence of the 3z1B01 heavy chain
136His Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly 1
5 10 15 Ala Ser Val Thr
Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp 20
25 30 Tyr Glu Ile His Trp Val Lys Gln Thr
Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Val Ile Asp Pro Glu Thr Gly Gly Thr Ala Tyr Asn
Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala 65
70 75 80 Tyr Met Glu Leu Arg
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln
Gly Thr Thr Leu Thr Val 100 105
110 Ser Ser 137114PRTArtificial SequenceAmino acid sequence of
the 3z1B02 heavy chain 137His Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu
Val Arg Pro Gly 1 5 10
15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp
20 25 30 Tyr Glu Ile
His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp 35
40 45 Ile Gly Val Ile Asp Pro Glu Thr
Gly Ala Thr Ala Tyr Asn Gln Lys 50 55
60 Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser
Ser Thr Ala 65 70 75
80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr
85 90 95 Cys Met Gly Tyr
Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100
105 110 Ser Ser 138114PRTArtificial
SequenceAmino acid sequence of the 3z1F04 heavy chain 138His Glu Val Gln
Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly 1 5
10 15 Ala Ser Val Thr Leu Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Asp 20 25
30 Tyr Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly Leu
Glu Trp 35 40 45
Ile Gly Val Ile Asp Pro Glu Thr Gly Ser Thr Ala Tyr Asn Gln Lys 50
55 60 Phe Lys Gly Lys Ala
Thr Leu Thr Ala Asp Lys Ala Ser Ser Thr Ala 65 70
75 80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu
Asp Ser Ala Val Tyr Tyr 85 90
95 Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr
Val 100 105 110 Ser
Ser 139114PRTArtificial SequenceAmino acid sequence of the 3z1E09 heavy
chain 139His Glu Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly
1 5 10 15 Ala Ser
Ala Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp 20
25 30 Tyr Glu Met His Trp Val Lys
Gln Thr Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Val Ile Asp Pro Glu Thr Gly Ser Thr Ala
Tyr Asn Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65
70 75 80 Tyr Met Glu
Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Met Gly Tyr Ala Asp Tyr Trp
Gly Gln Gly Thr Thr Leu Thr Val 100 105
110 Ser Ser 140114PRTArtificial SequenceAmino acid
sequence of the 3z1B08 heavy chain 140His Glu Val Gln Leu Gln Gln Ser Gly
Ala Glu Leu Val Arg Pro Gly 1 5 10
15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe
Thr Asp 20 25 30
Tyr Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp
35 40 45 Ile Gly Val Ile
Asp Pro Glu Thr Gly Asp Thr Ala Tyr Asn Gln Asn 50
55 60 Phe Thr Gly Lys Ala Thr Leu Thr
Ala Asp Lys Ser Ser Ser Thr Ala 65 70
75 80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser
Ala Val Tyr Tyr 85 90
95 Cys Met Gly Tyr Ala Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val
100 105 110 Ser Ser
141114PRTArtificial SequenceAmino acid sequence of the 3z1G08 heavy chain
141His Gln Val Gln Leu Lys Gln Ser Gly Ala Glu Leu Val Arg Pro Gly 1
5 10 15 Ala Ser Val Thr
Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp 20
25 30 Tyr Glu Val His Trp Val Lys Gln Thr
Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Val Ile Asp Pro Ala Thr Gly Asp Thr Ala Tyr Asn
Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65
70 75 80 Tyr Met Glu Val Ser
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln
Gly Thr Thr Leu Thr Val 100 105
110 Ser Ser 142114PRTArtificial SequenceAmino acid sequence of
the 3z1F07 heavy chain 142His Gln Ala Tyr Leu Gln Gln Ser Gly Ala Glu Leu
Val Arg Pro Gly 1 5 10
15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp
20 25 30 Tyr Glu Ile
His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp 35
40 45 Ile Gly Val Ile Asp Pro Glu Thr
Gly Asp Thr Ala Tyr Asn Gln Lys 50 55
60 Phe Lys Asp Lys Ala Thr Leu Thr Ala Asp Lys Ala Ser
Ser Thr Ala 65 70 75
80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr
85 90 95 Cys Met Gly Tyr
Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100
105 110 Ser Ser 143114PRTArtificial
SequenceAmino acid sequence of the 3z1E12 heavy chain 143His Gln Val Gln
Leu Gln Gln Ser Glu Ala Glu Leu Val Lys Pro Gly 1 5
10 15 Ala Ser Val Lys Leu Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Asp 20 25
30 Tyr Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly Leu
Glu Trp 35 40 45
Ile Gly Val Ile Asp Pro Glu Thr Gly Asp Thr Ala Tyr Asn Gln Lys 50
55 60 Phe Lys Gly Lys Ala
Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65 70
75 80 Tyr Met Glu Leu Ser Arg Leu Thr Ser Glu
Asp Ser Ala Val Tyr Tyr 85 90
95 Cys Met Gly His Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr
Val 100 105 110 Ser
Ser 144114PRTArtificial SequenceAmino acid sequence of the 3z1D03 heavy
chain 144His Glu Val Gln Leu Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly
1 5 10 15 Ala Ser
Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Ile Phe Thr Asp 20
25 30 Tyr Glu Ile His Trp Val Lys
Gln Thr Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Val Ile Asp Pro Glu Thr Gly Asn Thr Ala
Phe Asn Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Ile Ser Ser Ser Thr Ala 65
70 75 80 Tyr Met Glu
Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Met Gly Tyr Ser Asp Tyr Trp
Gly Gln Gly Thr Thr Leu Thr Val 100 105
110 Ser Ser 145114PRTArtificial SequenceAmino acid
sequence of the 3z1G12 heavy chain 145His Glu Val Gln Leu Gln Gln Ser Val
Ala Glu Leu Val Arg Pro Gly 1 5 10
15 Ala Ser Val Thr Val Ser Cys Lys Ala Ser Gly Tyr Ile Phe
Thr Asp 20 25 30
Tyr Glu Ile His Trp Val Lys Gln Thr Pro Ala His Gly Leu Glu Trp
35 40 45 Ile Gly Val Ile
Asp Pro Glu Thr Gly Asn Thr Ala Phe Asn Gln Lys 50
55 60 Phe Lys Gly Lys Ala Thr Leu Thr
Ala Asp Ile Ser Ser Ser Thr Ala 65 70
75 80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser
Ala Val Tyr Tyr 85 90
95 Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val
100 105 110 Ser Ser
146114PRTArtificial SequenceAmino acid sequence of the 3z1F10 heavy chain
146His Glu Val Gln Leu Gln Gln Ser Val Ala Glu Leu Val Arg Pro Gly 1
5 10 15 Ala Pro Val Thr
Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp 20
25 30 Tyr Glu Val His Trp Val Lys Gln Thr
Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Val Ile Asp Pro Glu Thr Gly Ala Thr Ala Tyr Asn
Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Ala Ala 65
70 75 80 Tyr Met Glu Leu Ser
Arg Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Met Ser Tyr Ser Asp Tyr Trp Gly Gln
Gly Thr Thr Leu Thr Val 100 105
110 Ser Ser 147114PRTArtificial SequenceAmino acid sequence of
the 3z1C03 heavy chain 147His Glu Val Gln Leu Gln Gln Ser Val Ala Glu Val
Val Arg Pro Gly 1 5 10
15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp
20 25 30 Tyr Glu Ile
His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp 35
40 45 Ile Gly Val Ile Asp Pro Glu Thr
Gly Val Thr Ala Tyr Asn Gln Arg 50 55
60 Phe Arg Asp Lys Ala Thr Leu Thr Thr Asp Lys Ser Ser
Ser Thr Ala 65 70 75
80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe
85 90 95 Cys Met Gly Tyr
Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val 100
105 110 Ser Ser 148114PRTArtificial
SequenceAmino acid sequence of the 3z1C03 heavy chain 148His Glu Val Gln
Leu Gln Gln Ser Val Ala Glu Val Val Arg Pro Gly 1 5
10 15 Ala Ser Val Thr Leu Ser Cys Lys Ala
Ser Gly Tyr Thr Phe Thr Asp 20 25
30 Tyr Glu Ile His Trp Val Lys Gln Thr Pro Val His Gly Leu
Glu Trp 35 40 45
Ile Gly Val Ile Asp Pro Glu Thr Gly Val Thr Ala Tyr Asn Gln Arg 50
55 60 Phe Arg Asp Lys Ala
Thr Leu Thr Thr Asp Lys Ser Ser Ser Thr Ala 65 70
75 80 Tyr Met Glu Leu Ser Ser Leu Thr Ser Glu
Asp Ser Ala Val Tyr Phe 85 90
95 Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr
Val 100 105 110 Ser
Ser 149114PRTArtificial SequenceAmino acid sequence of the 3z1G05 heavy
chain 149His Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly
1 5 10 15 Ala Ser
Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp 20
25 30 Tyr Glu Ile His Trp Val Lys
Gln Thr Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Val Leu Asp Pro Gly Thr Gly Arg Thr Ala
Tyr Asn Gln Lys 50 55 60
Phe Lys Asp Lys Ala Thr Leu Ser Ala Asp Lys Ser Ser Ser Thr Ala 65
70 75 80 Tyr Met Glu
Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Met Ser Tyr Ser Asp Tyr Trp
Gly Pro Gly Thr Thr Leu Thr Val 100 105
110 Ser Ser 150114PRTArtificial SequenceAmino acid
sequence of the 3z1B11 heavy chain 150His Glu Val Gln Leu Gln Gln Ser Val
Ala Glu Leu Val Arg Pro Gly 1 5 10
15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe
Thr Asp 20 25 30
Tyr Glu Met His Trp Val Lys Gln Thr Pro Val Arg Gly Leu Glu Trp
35 40 45 Ile Gly Val Ile
Asp Pro Ala Thr Gly Asp Thr Ala Tyr Asn Gln Lys 50
55 60 Phe Lys Gly Lys Ala Thr Leu Thr
Ala Asp Lys Ser Ser Ser Ala Ala 65 70
75 80 Phe Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser
Ala Val Tyr Tyr 85 90
95 Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr Thr Leu Thr Val
100 105 110 Ser Ser
151115PRTArtificial SequenceAmino acid sequence of the 3z1E06 heavy chain
151His Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly 1
5 10 15 Ala Ser Val Thr
Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ser Asp 20
25 30 Tyr Glu Met His Trp Val Lys Gln Thr
Pro Val His Gly Leu Glu Trp 35 40
45 Ile Gly Gly Ile Asp Pro Glu Thr Gly Asp Thr Val Tyr Asn
Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala 65
70 75 80 Tyr Met Glu Leu Ser
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Ile Ser Tyr Ala Met Asp Tyr Trp Gly
Gln Gly Thr Ser Val Thr 100 105
110 Val Ser Ser 115 152115PRTArtificial SequenceAmino
acid sequence of the 4z1A02 heavy chain 152His Gln Val Lys Leu Gln Gln
Ser Gly Thr Glu Leu Val Arg Pro Gly 1 5
10 15 Ala Ser Val Thr Leu Ser Cys Lys Ala Ser Gly
Tyr Lys Phe Thr Asp 20 25
30 Tyr Glu Met His Trp Val Lys Gln Thr Pro Val His Gly Leu Glu
Trp 35 40 45 Ile
Gly Gly Ile Asp Pro Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys 50
55 60 Phe Lys Gly Lys Ala Ile
Leu Thr Ala Asp Lys Ser Ser Thr Thr Ala 65 70
75 80 Tyr Met Glu Leu Arg Ser Leu Thr Ser Glu Asp
Ser Ala Val Tyr Tyr 85 90
95 Cys Ile Ser Tyr Ala Met Asp Tyr Trp Gly Gln Gly Thr Ser Val Thr
100 105 110 Val Ser
Ser 115 153117PRTArtificial SequenceAmino acid sequence of the
3z1E10 heavy chain 153His Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val
Lys Pro Gly 1 5 10 15
Ala Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Asp Thr Phe Thr Asp
20 25 30 Tyr Tyr Met Asn
Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu Trp 35
40 45 Ile Gly Asp Ile Asn Pro Asn Tyr Gly
Gly Ile Thr Tyr Asn Gln Lys 50 55
60 Phe Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser
Ser Thr Ala 65 70 75
80 Tyr Met Glu Leu Arg Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr
85 90 95 Cys Gln Ala Tyr
Tyr Arg Asn Ser Asp Tyr Trp Gly Gln Gly Thr Thr 100
105 110 Leu Thr Val Ser Ser 115
154116PRTArtificial SequenceAmino acid sequence of the 3z1G10 heavy
chain 154His Glu Ile Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly
1 5 10 15 Ala Ser
Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp 20
25 30 Asn Tyr Met Asn Trp Val Lys
Gln Ser His Gly Lys Ser Leu Glu Trp 35 40
45 Ile Gly Asp Ile Asn Pro Tyr Tyr Gly Thr Thr Thr
Tyr Asn Gln Lys 50 55 60
Phe Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Arg Thr Ala 65
70 75 80 Tyr Met Glu
Leu Arg Gly Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr 85
90 95 Cys Ala Arg Asp Asp Trp Phe Asp
Tyr Trp Gly Gln Gly Thr Leu Val 100 105
110 Thr Val Ser Ala 115 155118PRTArtificial
SequenceAmino acid sequence of the 3z1D01 heavy chain 155His Glu Val Gln
Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser 1 5
10 15 Gln Ser Leu Ser Leu Thr Cys Thr Val
Thr Gly Phe Ser Ile Thr Ser 20 25
30 Gly Tyr Gly Trp His Trp Ile Arg Gln Phe Pro Gly Asp Lys
Leu Glu 35 40 45
Trp Met Gly Tyr Ile Ser Phe Asn Gly Asp Tyr Asn Tyr Asn Pro Ser 50
55 60 Leu Lys Ser Arg Ile
Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe 65 70
75 80 Phe Leu Gln Leu Ser Ser Val Thr Thr Glu
Asp Thr Ala Thr Tyr Tyr 85 90
95 Cys Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu
Val Thr Val Ser Ala 115 156118PRTArtificial
SequenceAmino acid sequence of the 3z1C02 heavy chain 156His Asp Val Gln
Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser 1 5
10 15 Gln Ser Leu Ser Leu Thr Cys Thr Val
Thr Gly Phe Ser Ile Thr Ser 20 25
30 Gly Tyr Gly Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys
Leu Glu 35 40 45
Trp Met Gly Tyr Ile Ser Phe Asn Gly Asp Ser Asn Tyr Asn Pro Ser 50
55 60 Leu Lys Ser Arg Ile
Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe 65 70
75 80 Phe Leu Gln Leu Asn Ser Val Thr Ser Glu
Asp Thr Ala Thr Tyr Tyr 85 90
95 Cys Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr Trp Gly Gln Gly
Pro 100 105 110 Leu
Val Thr Val Ser Ala 115 157118PRTArtificial
SequenceAmino acid sequence of the 3z1C04 heavy chain 157His Glu Val Gln
Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser 1 5
10 15 Gln Ser Leu Ser Leu Thr Cys Thr Val
Thr Gly Phe Ser Ile Thr Ser 20 25
30 Gly Tyr Gly Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys
Leu Glu 35 40 45
Trp Met Gly Tyr Ile Asn Tyr Asp Gly His Asn Asp Tyr Asn Pro Ser 50
55 60 Leu Lys Ser Arg Ile
Ser Ile Thr Gln Asp Thr Ser Lys Asn Gln Phe 65 70
75 80 Phe Leu Gln Leu Asn Ser Val Thr Thr Glu
Asp Thr Ala Thr Tyr Tyr 85 90
95 Cys Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr Trp Gly Gln Gly
Thr 100 105 110 Leu
Val Thr Val Ser Ala 115 15816PRTArtificial
SequenceAmino acid sequence of the 3z1A02 light chain CDR1 158Lys
Ser Ser Gln Ser Leu Leu His Ser Asp Gly Lys Thr Tyr Leu Asn 1
5 10 15 1597PRTArtificial
SequenceAmino acid sequence of the 3z1A02 light chain CDR2 159Leu
Val Ser Lys Leu Asp Ser 1 5 1609PRTArtificial
SequenceAmino acid sequence of the 3z1A02 light chain CDR3 160Trp
Gln Gly Thr His Phe Pro Arg Thr 1 5
16110PRTArtificial SequenceAmino acid sequence of the 3z1A02 heavy chain
CDR1 161Gly Tyr Thr Phe Thr Asp Tyr Asn Met His 1 5
10 16210PRTArtificial SequenceAmino acid sequence of the
3z1A02 heavy chain CDR2 162Tyr Ile Asn Pro Tyr Asn Asp Val Thr Glu 1
5 10 1637PRTArtificial SequenceAmino
acid sequence of the 3z1A02 heavy chain CDR3 163Ala Trp Phe Gly Leu
Arg Gln 1 5 16416PRTArtificial SequenceAmino acid
sequence of the 3z1E10 light chain CDR1 164Arg Ser Ser Lys Ser Leu
Leu His Ser Asn Gly Asn Thr Tyr Leu Tyr 1 5
10 15 1657PRTArtificial SequenceAmino acid
sequence of the 3z1E10 light chain CDR2 165Arg Met Ser Asn Leu Ala
Ser 1 5 1669PRTArtificial SequenceAmino acid
sequence of the 3z1E10 light chain CDR3 166Met Gln His Leu Glu Tyr
Pro Tyr Thr 1 5 16710PRTArtificial
SequenceAmino acid sequence of the 3z1E10 heavy chain CDR1 167Gly
Asp Thr Phe Thr Asp Tyr Tyr Met Asn 1 5
10 16810PRTArtificial SequenceAmino acid sequence of the 3z1E10 heavy
chain CDR2 168Asp Ile Asn Pro Asn Tyr Gly Gly Ile Thr 1
5 10 1699PRTArtificial SequenceAmino acid sequence of
the 3z1E10 heavy chain CDR3 169Gln Ala Tyr Tyr Arg Asn Ser Asp Tyr 1
5 17011PRTArtificial SequenceAmino acid
sequence of the 3z1G12 light chain CDR1 170Lys Ala Ser Gln Asp Val
Gly Thr Ala Val Ala 1 5 10
1717PRTArtificial SequenceAmino acid sequence of the 3z1G12 light chain
CDR2 171Trp Thr Ser Thr Arg His Thr 1 5
1729PRTArtificial SequenceAmino acid sequence of the 3z1G12 light chain
CDR3 172Gln Gln His Tyr Ser Ile Pro Leu Thr 1 5
17310PRTArtificial SequenceAmino acid sequence of the 3z1G12
heavy chain CDR1 173Gly Tyr Ile Phe Thr Asp Tyr Glu Ile His 1
5 10 17410PRTArtificial SequenceAmino acid
sequence of the 3z1G12 heavy chain CDR2 174Val Ile Asp Pro Glu Thr
Gly Asn Thr Ala 1 5 10 1756PRTArtificial
SequenceAmino acid sequence of the 3z1G12 heavy chain CDR3 175Met
Gly Tyr Ser Asp Tyr 1 5 176240PRTArtificial
SequenceHumanized 3D3 antibody light chain 176Met Val Leu Gln Thr Gln Val
Phe Ile Ser Leu Leu Leu Trp Ile Ser 1 5
10 15 Gly Ala Tyr Gly Asp Ile Val Met Thr Gln Ser
Pro Asp Ser Leu Ala 20 25
30 Val Ser Leu Gly Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln
Ser 35 40 45 Leu
Leu Asn Ser Asn Phe Gln Lys Asn Phe Leu Ala Trp Tyr Gln Gln 50
55 60 Lys Pro Gly Gln Pro Pro
Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg 65 70
75 80 Glu Ser Ser Val Pro Asp Arg Phe Ser Gly Ser
Gly Ser Gly Thr Asp 85 90
95 Phe Thr Leu Thr Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr
100 105 110 Tyr Cys
Gln Gln His Tyr Ser Thr Pro Leu Thr Phe Gly Gln Gly Thr 115
120 125 Lys Leu Glu Ile Lys Arg Thr
Val Ala Ala Pro Ser Val Phe Ile Phe 130 135
140 Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
Ser Val Val Cys 145 150 155
160 Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val
165 170 175 Asp Asn Ala
Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln 180
185 190 Asp Ser Lys Asp Ser Thr Tyr Ser
Leu Ser Ser Thr Leu Thr Leu Ser 195 200
205 Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu
Val Thr His 210 215 220
Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225
230 235 240
177462PRTArtificial SequenceHumanized 3D3 antibody heavy chain 177Met Asp
Trp Thr Trp Arg Ile Leu Phe Leu Val Ala Ala Ala Thr Gly 1 5
10 15 Thr His Ala Glu Val Gln Leu
Val Gln Ser Gly Ala Glu Val Lys Lys 20 25
30 Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser
Gly Tyr Ile Phe 35 40 45
Thr Asp Tyr Glu Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
50 55 60 Glu Trp Met
Gly Val Ile Asp Pro Glu Thr Gly Asn Thr Ala Phe Asn 65
70 75 80 Gln Lys Phe Lys Gly Arg Val
Thr Ile Thr Ala Asp Thr Ser Thr Ser 85
90 95 Thr Ala Tyr Met Glu Leu Ser Ser Leu Thr Ser
Glu Asp Thr Ala Val 100 105
110 Tyr Tyr Cys Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly Thr Leu
Val 115 120 125 Thr
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 130
135 140 Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 145 150
155 160 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser Gly 165 170
175 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
180 185 190 Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 195
200 205 Gly Thr Gln Thr Tyr Ile Cys
Asn Val Asn His Lys Pro Ser Asn Thr 210 215
220 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
Lys Thr His Thr 225 230 235
240 Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
245 250 255 Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 260
265 270 Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro Glu Val 275 280
285 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
Ala Lys Thr 290 295 300
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 305
310 315 320 Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 325
330 335 Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile Glu Lys Thr Ile Ser 340 345
350 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
Pro Pro 355 360 365
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 370
375 380 Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 385 390
395 400 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val Leu Asp Ser Asp 405 410
415 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
Trp 420 425 430 Gln
Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 435
440 445 Asn His Tyr Thr Gln Lys
Ser Leu Ser Leu Ser Pro Gly Lys 450 455
460 178113PRTArtificial SequenceHumanized 3D3 antibody light
chain variable region 178Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu
Ala Val Ser Leu Gly 1 5 10
15 Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
20 25 30 Asn Phe
Gln Lys Asn Phe Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35
40 45 Pro Pro Lys Leu Leu Ile Tyr
Phe Ala Ser Thr Arg Glu Ser Ser Val 50 55
60 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr 65 70 75
80 Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln
85 90 95 His Tyr Ser
Thr Pro Leu Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 100
105 110 Lys 179113PRTArtificial
SequenceHumanized 3D3 antibody heavy chain variable region 179Glu
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1
5 10 15 Ser Val Lys Val Ser Cys
Lys Ala Ser Gly Tyr Ile Phe Thr Asp Tyr 20
25 30 Glu Ile His Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Met 35 40
45 Gly Val Ile Asp Pro Glu Thr Gly Asn Thr Ala Phe Asn Gln
Lys Phe 50 55 60
Lys Gly Arg Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Ser Ser
Leu Thr Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly
Thr Leu Val Thr Val Ser 100 105
110 Ser 180234PRTArtificial SequenceHumanized 3C4 antibody
light chain 180Met Val Leu Gln Thr Gln Val Phe Ile Ser Leu Leu Leu Trp
Ile Ser 1 5 10 15
Gly Ala Tyr Gly Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser
20 25 30 Ala Ser Val Gly Asp
Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp 35
40 45 Ile His Asn Phe Leu Asn Trp Phe Gln
Gln Lys Pro Gly Lys Ala Pro 50 55
60 Lys Thr Leu Ile Phe Arg Ala Asn Arg Leu Val Asp Gly
Val Pro Ser 65 70 75
80 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser
85 90 95 Ser Leu Gln Pro
Glu Asp Phe Ala Thr Tyr Ser Cys Leu Gln Tyr Asp 100
105 110 Glu Ile Pro Leu Thr Phe Gly Gln Gly
Thr Lys Leu Glu Ile Lys Arg 115 120
125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp
Glu Gln 130 135 140
Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr 145
150 155 160 Pro Arg Glu Ala Lys
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165
170 175 Gly Asn Ser Gln Glu Ser Val Thr Glu Gln
Asp Ser Lys Asp Ser Thr 180 185
190 Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
Lys 195 200 205 His
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210
215 220 Val Thr Lys Ser Phe Asn
Arg Gly Glu Cys 225 230
181466PRTArtificial SequenceHumanized 3C4 antibody heavy chain 181Met Asp
Trp Thr Trp Arg Ile Leu Phe Leu Val Ala Ala Ala Thr Gly 1 5
10 15 Thr His Ala Glu Val Gln Leu
Gln Glu Ser Gly Pro Gly Leu Val Lys 20 25
30 Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser
Gly Phe Ser Ile 35 40 45
Thr Ser Gly Tyr Gly Trp His Trp Ile Arg Gln His Pro Gly Lys Gly
50 55 60 Leu Glu Trp
Ile Gly Tyr Ile Asn Tyr Asp Gly His Asn Asp Tyr Asn 65
70 75 80 Pro Ser Leu Lys Ser Arg Val
Thr Ile Ser Gln Asp Thr Ser Lys Asn 85
90 95 Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala
Ala Asp Thr Ala Val 100 105
110 Tyr Tyr Cys Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr Trp Gly
Gln 115 120 125 Gly
Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 130
135 140 Phe Pro Leu Ala Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 145 150
155 160 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser 165 170
175 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
180 185 190 Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 195
200 205 Ser Ser Ser Leu Gly Thr Gln
Thr Tyr Ile Cys Asn Val Asn His Lys 210 215
220 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys Asp 225 230 235
240 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
245 250 255 Pro Ser Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 260
265 270 Ser Arg Thr Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His Glu 275 280
285 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His 290 295 300
Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 305
310 315 320 Val Val Ser Val
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 325
330 335 Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu 340 345
350 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
Val Tyr 355 360 365
Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 370
375 380 Thr Cys Leu Val Lys
Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 385 390
395 400 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
Lys Thr Thr Pro Pro Val 405 410
415 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Asp 420 425 430 Lys
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 435
440 445 Glu Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 450 455
460 Gly Lys 465 182107PRTArtificial
SequenceHumanized 3C4 antibody light chain variable region 182Asp
Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1
5 10 15 Asp Arg Val Thr Ile Thr
Cys Lys Ala Ser Gln Asp Ile His Asn Phe 20
25 30 Leu Asn Trp Phe Gln Gln Lys Pro Gly Lys
Ala Pro Lys Thr Leu Ile 35 40
45 Phe Arg Ala Asn Arg Leu Val Asp Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro 65
70 75 80 Glu Asp Phe Ala Thr
Tyr Ser Cys Leu Gln Tyr Asp Glu Ile Pro Leu 85
90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys 100 105 183116PRTArtificial
SequenceHumanized 3C4 antibody heavy chain variable region 183Glu
Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln 1
5 10 15 Thr Leu Ser Leu Thr Cys
Thr Val Ser Gly Phe Ser Ile Thr Ser Gly 20
25 30 Tyr Gly Trp His Trp Ile Arg Gln His Pro
Gly Lys Gly Leu Glu Trp 35 40
45 Ile Gly Tyr Ile Asn Tyr Asp Gly His Asn Asp Tyr Asn Pro
Ser Leu 50 55 60
Lys Ser Arg Val Thr Ile Ser Gln Asp Thr Ser Lys Asn Gln Phe Ser 65
70 75 80 Leu Lys Leu Ser Ser
Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr
Trp Gly Gln Gly Thr Leu 100 105
110 Val Thr Val Ser 115 18417PRTArtificial
SequenceCDRL1 corresponding to residues 24-40 of SEQ ID NO.111
184Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys Asn Tyr Leu 1
5 10 15 Ala
18517PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of SEQ
ID NO.112 185Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys Asn
Tyr Leu 1 5 10 15
Ala 18617PRTArtificial SequenceCDRL1 corresponding to resid3es 24-40 of
SEQ ID NO.113 186Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 18717PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.114 187Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 18817PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.115 188Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 18917PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.116 189Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19017PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.117 190Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19117PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.118 191Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19217PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.123 192Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19317PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.125 193Lys Ser Ser Gln Ser Leu Leu Asn Ser Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19417PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.119 194Lys Ser Ser Gln Ser Leu Leu Asn Arg Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19517PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.120 195Lys Ser Ser Gln Ser Leu Leu Asn Asn Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19617PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.121 196Lys Ser Ser Gln Ser Leu Leu Asn Thr Ser Asn Gln Leu
Asn Tyr Leu 1 5 10 15
Ala 19717PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.122 197Lys Ser Ser Gln Ser Leu Leu Asn Thr Ser Asn Gln Lys
Asn Tyr Leu 1 5 10 15
Ala 19817PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.110 198Lys Ser Ser Gln Ser Leu Leu Asn Ser Asn Asn Gln Leu
Asn Tyr Leu 1 5 10 15
Ala 19917PRTArtificial SequenceCDRL1 corresponding to residues 24-40 of
SEQ ID NO.124 199Lys Ser Ser Gln Ser Leu Leu Asn Ser Asn Phe Gln Lys
Asn Phe Leu 1 5 10 15
Ala 20016PRTArtificial SequenceCDRL1 corresponding to residues 24-39 of
SEQ ID NO.105 200Lys Ser Ser Gln Ser Leu Leu His Ser Asp Gly Lys Thr
Tyr Leu Asn 1 5 10 15
20116PRTArtificial SequenceCDRL1 corresponding to residues 24-39 of SEQ
ID NO.107 201Lys Ser Ser Gln Ser Leu Leu His Ser Asp Gly Lys Thr Tyr
Leu Asn 1 5 10 15
20216PRTArtificial SequenceCDRL1 corresponding to residues 24-39 of SEQ
ID NO.106 202Lys Ser Ser Gln Ser Leu Leu Tyr Ser Asp Gly Lys Thr Tyr
Leu Asn 1 5 10 15
20316PRTArtificial SequenceCDRL1 corresponding to residues 24-39 of SEQ
ID NO.108 203Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Asn Thr Tyr
Leu Glu 1 5 10 15
20416PRTArtificial SequenceCDRL1 corresponding to residues 24-39 of SEQ
ID NO.109 204Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Tyr
Leu Tyr 1 5 10 15
20511PRTArtificial SequenceCDRL1 corresponding to residues 24-34 of SEQ
ID NO.127 205Lys Ala Ser Gln Asp Ile His Asn Phe Leu Asn 1
5 10 20611PRTArtificial SequenceCDRL1
corresponding to residues 24-34 of SEQ ID NO.131 206Lys Ala Ser Gln
Asp Ile His Arg Phe Leu Asn 1 5 10
20711PRTArtificial SequenceCDRL1 corresponding to residues 24-34 of SEQ
ID NO.129 207Lys Ala Ser Gln Asp Ile His Asn Tyr Leu Asn 1
5 10 20811PRTArtificial SequenceCDRL1
corresponding to residues 24-34 of SEQ ID NO.130 208Lys Ala Ser Gln
Asp Ile His Asn Tyr Leu Asn 1 5 10
20911PRTArtificial SequenceCDRL1 corresponding to residues 24-34 of SEQ
ID NO.128 209Lys Ala Ser Gln Asp Ile His Thr Tyr Leu Asn 1
5 10 21011PRTArtificial SequenceCDRL1
corresponding to residues 24-34 of SEQ ID NO.126 210Lys Ala Ser Gln
Asp Val Gly Thr Ala Val Ala 1 5 10
2117PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.111 211Phe Ala Ser Thr Arg Glu Ser 1 5
2127PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.112 212Phe Ala Ser Thr Arg Glu Ser 1 5
2137PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.121 213Phe Ala Ser Thr Thr Glu Ser 1 5
2147PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.113 214Phe Ala Ser Thr Arg Glu Ser 1 5
2157PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.114 215Phe Ala Ser Thr Arg Glu Ser 1 5
2167PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.125 216Phe Gly Ser Thr Arg Glu Ser 1 5
2177PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.118 217Phe Gly Ser Thr Arg Glu Ser 1 5
2187PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.115 218Phe Ala Ser Thr Arg Glu Ser 1 5
2197PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.116 219Phe Ala Ser Thr Arg Glu Ser 1 5
2207PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.117 220Phe Ala Ser Thr Arg Glu Ser 1 5
2217PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.119 221Phe Ala Ser Thr Arg Glu Ser 1 5
2227PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.120 222Phe Ala Ser Thr Arg Glu Ser 1 5
2237PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.123 223Phe Ala Ser Thr Arg Glu Ser 1 5
2247PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.124 224Phe Ala Ser Thr Arg Glu Ser 1 5
2257PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.110 225Phe Ala Ser Thr Arg Lys Ser 1 5
2267PRTArtificial SequenceCDRL2 corresponding to residues 56-62 of SEQ ID
NO.122 226Phe Ala Ser Thr Arg Ala Ser 1 5
2277PRTArtificial SequenceCDRL2 corresponding to residues 55-61 of SEQ ID
NO.105 227Leu Val Ser Lys Leu Asp Ser 1 5
2287PRTArtificial SequenceCDRL2 corresponding to residues 55-61 of SEQ ID
NO.106 228Leu Val Ser Lys Leu Asp Ser 1 5
2297PRTArtificial SequenceCDRL2 corresponding to residues 55-61 of SEQ ID
NO.107 229Leu Val Ser Lys Leu Asp Ser 1 5
2307PRTArtificial SequenceCDRL2 corresponding to residues 55-61 of SEQ ID
NO.108 230Lys Val Ser Asn Arg Phe Ser 1 5
2317PRTArtificial SequenceCDRL2 corresponding to residues 50-56 of SEQ ID
NO.127 231Arg Ala Asn Arg Leu Val Asp 1 5
2327PRTArtificial SequenceCDRL2 corresponding to residues 50-56 of SEQ ID
NO.128 232Arg Ala Asn Arg Leu Val Asp 1 5
2337PRTArtificial SequenceCDRL2 corresponding to residues 50-56 of SEQ ID
NO.131 233His Ala Asn Arg Leu Val Asp 1 5
2347PRTArtificial SequenceCDRL2 corresponding to residues 50-56 of SEQ ID
NO.129 234Arg Ala Asn Arg Leu Val Ala 1 5
2357PRTArtificial SequenceCDRL2 corresponding to residues 50-56 of SEQ ID
NO.130 235Arg Ala Asn Arg Leu Val Ala 1 5
2369PRTArtificial SequenceCDRL3 corresponding to residues 95-103 of SEQ
ID NO.114 236Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
2379PRTArtificial SequenceCDRL3 corresponding to residues
95-103 of SEQ ID NO.115 237Gln Gln His Tyr Ser Thr Pro Leu Thr 1
5 2389PRTArtificial SequenceCDRL3
corresponding to residues 95-103 of SEQ ID NO.116 238Gln Gln His Tyr
Ser Thr Pro Leu Thr 1 5 2399PRTArtificial
SequenceCDRL3 corresponding to residues 95-103 of SEQ ID NO.117
239Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
2409PRTArtificial SequenceCDRL3 corresponding to residues 95-103 of SEQ
ID NO.118 240Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
2419PRTArtificial SequenceCDRL3 corresponding to residues
95-103 of SEQ ID NO.120 241Gln Gln His Tyr Ser Thr Pro Leu Thr 1
5 2429PRTArtificial SequenceCDRL3
corresponding to residues 95-103 of SEQ ID NO.121 242Gln Gln His Tyr
Ser Thr Pro Leu Thr 1 5 2439PRTArtificial
SequenceCDRL3 corresponding to residues 95-103 of SEQ ID NO.122
243Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
2449PRTArtificial SequenceCDRL3 corresponding to residues 95-103 of SEQ
ID NO.123 244Gln Gln His Tyr Ser Thr Pro Leu Thr 1 5
2459PRTArtificial SequenceCDRL3 corresponding to residues
95-103 of SEQ ID NO.124 245Gln Gln His Tyr Ser Thr Pro Leu Thr 1
5 2469PRTArtificial SequenceCDRL3
corresponding to residues 95-103 of SEQ ID NO.125 246Gln Gln His Tyr
Ser Thr Pro Leu Thr 1 5 2479PRTArtificial
SequenceCDRL3 corresponding to residues 95-103 of SEQ ID NO.111
247Gln Gln His Tyr Ser Ile Pro Leu Thr 1 5
2489PRTArtificial SequenceCDRL3 corresponding to residues 95-103 of SEQ
ID NO.112 248Gln Gln His Tyr Ser Ile Pro Leu Thr 1 5
2499PRTArtificial SequenceCDRL3 corresponding to residues
95-103 of SEQ ID NO.113 249Gln Gln His Tyr Ser Ile Pro Leu Thr 1
5 2509PRTArtificial SequenceCDRL3
corresponding to residues 95-103 of SEQ ID NO.119 250Gln Gln His Tyr
Ser Ile Pro Leu Thr 1 5 2519PRTArtificial
SequenceCDRL3 corresponding to residues 89-97 of SEQ ID NO.126
251Gln Gln His Tyr Ser Ile Pro Leu Thr 1 5
2529PRTArtificial SequenceCDRL3 corresponding to residues 95-103 of SEQ
ID NO.110 252Gln Gln His Phe Asn Thr Pro Leu Thr 1 5
2539PRTArtificial SequenceCDRL3 corresponding to residues
89-97 of SEQ ID NO.129 253Leu Gln Tyr Asp Ala Phe Pro Leu Thr 1
5 2549PRTArtificial SequenceCDRL3
corresponding to residues 89-97 of SEQ ID NO.130 254Leu Gln Tyr Asp
Ala Phe Pro Leu Thr 1 5 2559PRTArtificial
SequenceCDRL3 corresponding to residues 89-97 of SEQ ID NO.131
255Leu Gln Tyr Asp Ala Phe Pro Leu Thr 1 5
2569PRTArtificial SequenceCDRL3 corresponding to residues 89-97 of SEQ
ID NO.128 256Leu Gln Tyr Asp Glu Phe Pro Leu Thr 1 5
2579PRTArtificial SequenceCDRL3 corresponding to residues
89-97 of SEQ ID NO.127 257Leu Gln Tyr Asp Glu Ile Pro Leu Thr 1
5 2589PRTArtificial SequenceCDRL3
corresponding to residues 94-102 of SEQ ID NO.105 258Trp Gln Gly Thr
His Phe Pro Arg Thr 1 5 2599PRTArtificial
SequenceCDRL3 corresponding to residues 94-102 of SEQ ID NO.106
259Trp Gln Gly Thr His Phe Pro Arg Thr 1 5
2609PRTArtificial SequenceCDRL3 corresponding to residues 94-102 of SEQ
ID NO.107 260Trp Gln Gly Thr His Phe Pro Arg Thr 1 5
2619PRTArtificial SequenceCDRL3 corresponding to residues
94-102 of SEQ ID NO.108 261Phe Gln Gly Ser His Val Pro Leu Thr 1
5 26210PRTArtificial SequenceCDRH1
corresponding to residues 27-36 of SEQ ID NO.136 262Gly Tyr Thr Phe
Thr Asp Tyr Glu Ile His 1 5 10
26310PRTArtificial SequenceCDRH1 corresponding to residues 27-36 of SEQ
ID NO.137 263Gly Tyr Thr Phe Thr Asp Tyr Glu Ile His 1
5 10 26410PRTArtificial SequenceCDRH1 corresponding to
residues 27-36 of SEQ ID NO.138 264Gly Tyr Thr Phe Thr Asp Tyr Glu
Ile His 1 5 10 26510PRTArtificial
SequenceCDRH1 corresponding to residues 27-36 of SEQ ID NO.140
265Gly Tyr Thr Phe Thr Asp Tyr Glu Ile His 1 5
10 26610PRTArtificial SequenceCDRH1 corresponding to residues 27-36
of SEQ ID NO.142 266Gly Tyr Thr Phe Thr Asp Tyr Glu Ile His 1
5 10 26710PRTArtificial SequenceCDRH1
corresponding to residues 27-36 of SEQ ID NO.143 267Gly Tyr Thr Phe
Thr Asp Tyr Glu Ile His 1 5 10
26810PRTArtificial SequenceCDRH1 corresponding to residues 27-36 of SEQ
ID NO.147 268Gly Tyr Thr Phe Thr Asp Tyr Glu Ile His 1
5 10 26910PRTArtificial SequenceCDRH1 corresponding to
residues 27-36 of SEQ ID NO.148 269Gly Tyr Thr Phe Thr Asp Tyr Glu
Ile His 1 5 10 27010PRTArtificial
SequenceCDRH1 corresponding to residues 27-36 of SEQ ID NO.149
270Gly Tyr Thr Phe Thr Asp Tyr Glu Ile His 1 5
10 27110PRTArtificial SequenceCDRH1 corresponding to residues 27-36
of SEQ ID NO.144 271Gly Tyr Ile Phe Thr Asp Tyr Glu Ile His 1
5 10 27210PRTArtificial SequenceCDRH1
corresponding to residues 27-36 of SEQ ID NO.145 272Gly Tyr Ile Phe
Thr Asp Tyr Glu Ile His 1 5 10
27310PRTArtificial SequenceCDRH1 corresponding to residues 27-36 of SEQ
ID NO.135 273Gly Tyr Ile Phe Thr Asp Tyr Glu Val His 1
5 10 27410PRTArtificial SequenceCDRH1 corresponding to
residues 27-36 of SEQ ID NO.141 274Gly Tyr Thr Phe Thr Asp Tyr Glu
Val His 1 5 10 27510PRTArtificial
SequenceCDRH1 corresponding to residues 27-36 of SEQ ID NO.146
275Gly Tyr Thr Phe Thr Asp Tyr Glu Val His 1 5
10 27610PRTArtificial SequenceCDRH1 corresponding to residues 27-36
of SEQ ID NO.139 276Gly Tyr Thr Phe Thr Asp Tyr Glu Met His 1
5 10 27710PRTArtificial SequenceCDRH1
corresponding to residues 27-36 of SEQ ID NO.151 277Gly Tyr Thr Phe
Ser Asp Tyr Glu Met His 1 5 10
27810PRTArtificial SequenceCDRH1 corresponding to residues 27-36 of SEQ
ID NO.150 278Gly Tyr Thr Phe Thr Asp Tyr Glu Met His 1
5 10 27910PRTArtificial SequenceCDRH1 corresponding to
residues 27-36 of SEQ ID NO.152 279Gly Tyr Lys Phe Thr Asp Tyr Glu
Met His 1 5 10 28010PRTArtificial
SequenceCDRH1 corresponding to residues 27-36 of SEQ ID NO.132
280Gly Tyr Thr Phe Thr Asp Tyr Asn Met His 1 5
10 28110PRTArtificial SequenceCDRH1 corresponding to residues 27-36
of SEQ ID NO.133 281Gly Tyr Ile Phe Thr Glu Tyr Asn Ile His 1
5 10 28210PRTArtificial SequenceCDRH1
corresponding to residues 27-36 of SEQ ID NO.134 282Gly Tyr Thr Phe
Thr Glu Tyr Asn Met His 1 5 10
28311PRTArtificial SequenceCDRH1 corresponding to residues 27-37 of SEQ
ID NO.155 283Gly Phe Ser Ile Thr Ser Gly Tyr Gly Trp His 1
5 10 28411PRTArtificial SequenceCDRH1
corresponding to residues 27-37 of SEQ ID NO.156 284Gly Phe Ser Ile
Thr Ser Gly Tyr Gly Trp His 1 5 10
28510PRTArtificial SequenceCDRH2 corresponding to residues 51-60 of SEQ
ID NO.141 285Val Ile Asp Pro Ala Thr Gly Asp Thr Ala 1
5 10 28610PRTArtificial SequenceCDRH2 corresponding to
residues 51-60 of SEQ ID NO.150 286Val Ile Asp Pro Ala Thr Gly Asp
Thr Ala 1 5 10 28710PRTArtificial
SequenceCDRH2 corresponding to residues 51-60 of SEQ ID NO.135
287Val Ile Asp Pro Glu Thr Gly Asp Thr Ala 1 5
10 28810PRTArtificial SequenceCDRH2 corresponding to residues 51-60
of SEQ ID NO.142 288Val Ile Asp Pro Glu Thr Gly Asp Thr Ala 1
5 10 28910PRTArtificial SequenceCDRH2
corresponding to residues 51-60 of SEQ ID NO.143 289Val Ile Asp Pro
Glu Thr Gly Asp Thr Ala 1 5 10
29010PRTArtificial SequenceCDRH2 corresponding to residues 51-60 of SEQ
ID NO.140 290Val Ile Asp Pro Glu Thr Gly Asp Thr Ala 1
5 10 29110PRTArtificial SequenceCDRH2 corresponding to
residues 51-60 of SEQ ID NO.147 291Val Ile Asp Pro Glu Thr Gly Val
Thr Ala 1 5 10 29210PRTArtificial
SequenceCDRH2 corresponding to residues 51-60 of SEQ ID NO.148
292Val Ile Asp Pro Glu Thr Gly Val Thr Ala 1 5
10 29310PRTArtificial SequenceCDRH2 corresponding to residues 51-60
of SEQ ID NO.144 293Val Ile Asp Pro Glu Thr Gly Asn Thr Ala 1
5 10 29410PRTArtificial SequenceCDRH2
corresponding to residues 51-60 of SEQ ID NO.145 294Val Ile Asp Pro
Glu Thr Gly Asn Thr Ala 1 5 10
29510PRTArtificial SequenceCDRH2 corresponding to residues 51-60 of SEQ
ID NO.138 295Val Ile Asp Pro Glu Thr Gly Ser Thr Ala 1
5 10 29610PRTArtificial SequenceCDRH2 corresponding to
residues 51-60 of SEQ ID NO.139 296Val Ile Asp Pro Glu Thr Gly Ser
Thr Ala 1 5 10 29710PRTArtificial
SequenceCDRH2 corresponding to residues 51-60 of SEQ ID NO.137
297Val Ile Asp Pro Glu Thr Gly Ala Thr Ala 1 5
10 29810PRTArtificial SequenceCDRH2 corresponding to residues 51-60
of SEQ ID NO.146 298Val Ile Asp Pro Glu Thr Gly Ala Thr Ala 1
5 10 29910PRTArtificial SequenceCDRH2
corresponding to residues 51-60 of SEQ ID NO.151 299Gly Ile Asp Pro
Glu Thr Gly Asp Thr Val 1 5 10
30010PRTArtificial SequenceCDRH2 corresponding to residues 51-60 of SEQ
ID NO.152 300Gly Ile Asp Pro Glu Thr Gly Gly Thr Ala 1
5 10 30110PRTArtificial SequenceCDRH2 corresponding to
residues 51-60 of SEQ ID NO.136 301Val Ile Asp Pro Glu Thr Gly Gly
Thr Ala 1 5 10 30210PRTArtificial
SequenceCDRH2 corresponding to residues 51-60 of SEQ ID NO.149
302Val Leu Asp Pro Gly Thr Gly Arg Thr Ala 1 5
10 3039PRTArtificial SequenceCDRH2 corresponding to residues 52-60
of SEQ ID NO.155 303Tyr Ile Ser Phe Asn Gly Asp Tyr Asn 1
5 3049PRTArtificial SequenceCDRH2 corresponding to
residues 52-60 of SEQ ID NO.156 304Tyr Ile Ser Phe Asn Gly Asp Ser
Asn 1 5 3059PRTArtificial SequenceCDRH2
corresponding to residues 52-60 of SEQ ID NO.157 305Tyr Ile Asn Tyr
Asp Gly His Asn Asp 1 5
30610PRTArtificial SequenceCDRH2 corresponding to residues 51-60 of SEQ
ID NO.133 306Asn Ile Asn Pro Tyr Asn Asp Val Thr Glu 1
5 10 30710PRTArtificial SequenceCDRH2 corresponding to
residues 51-60 of SEQ ID NO.134 307Asn Ile Asn Pro Tyr Asn Asn Val
Thr Glu 1 5 10 30810PRTArtificial
SequenceCDRH2 corresponding to residues 51-60 of SEQ ID NO.132
308Tyr Ile Asn Pro Tyr Asn Asp Val Thr Glu 1 5
10 30910PRTArtificial SequenceCDRH2 corresponding to residues 51-60
of SEQ ID NO.153 309Asp Ile Asn Pro Asn Tyr Gly Gly Ile Thr 1
5 10 31010PRTArtificial SequenceCDRH2
corresponding to residues 51-60 of SEQ ID NO.154 310Asp Ile Asn Pro
Tyr Tyr Gly Thr Thr Thr 1 5 10
3116PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.146 311Met Ser Tyr Ser Asp Tyr 1 5
3126PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.149 312Met Ser Tyr Ser Asp Tyr 1 5
3136PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.136 313Met Gly Tyr Ser Asp Tyr 1 5
3146PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.137 314Met Gly Tyr Ser Asp Tyr 1 5
3156PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.138 315Met Gly Tyr Ser Asp Tyr 1 5
3166PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.141 316Met Gly Tyr Ser Asp Tyr 1 5
3176PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.142 317Met Gly Tyr Ser Asp Tyr 1 5
3186PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.143 318Met Gly His Ser Asp Tyr 1 5
3196PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.144 319Met Gly Tyr Ser Asp Tyr 1 5
3206PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.145 320Met Gly Tyr Ser Asp Tyr 1 5
3216PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.147 321Met Gly Tyr Ser Asp Tyr 1 5
3226PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.148 322Met Gly Tyr Ser Asp Tyr 1 5
3236PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.150 323Met Gly Tyr Ser Asp Tyr 1 5
3246PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.139 324Met Gly Tyr Ala Asp Tyr 1 5
3256PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.140 325Met Gly Tyr Ala Asp Tyr 1 5
3267PRTArtificial SequenceCDRH3 corresponding to residues 98-104 of SEQ
ID NO.151 326Ile Ser Tyr Ala Met Asp Tyr 1 5
3277PRTArtificial SequenceCDRH3 corresponding to residues 98-104 of SEQ
ID NO.152 327Ile Ser Tyr Ala Met Asp Tyr 1 5
3286PRTArtificial SequenceCDRH3 corresponding to residues 98-103 of SEQ
ID NO.135 328Ile Gly Tyr Ala Asp Tyr 1 5
3297PRTArtificial SequenceCDRH3 corresponding to residues 98-104 of SEQ
ID NO.133 329Ala Arg Trp Gly Leu Arg Asn 1 5
3307PRTArtificial SequenceCDRH3 corresponding to residues 98-104 of SEQ
ID NO.134 330Ala Arg Trp Gly Leu Arg Asn 1 5
3317PRTArtificial SequenceCDRH3 corresponding to residues 98-104 of SEQ
ID NO.132 331Ala Trp Phe Gly Leu Arg Gln 1 5
33210PRTArtificial SequenceCDRH3 corresponding to residues 98-107 of SEQ
ID NO.155 332Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr 1
5 10 33310PRTArtificial SequenceCDRH3 corresponding to
residues 98-107 of SEQ ID NO.156 333Ala Ser Ser Tyr Asp Gly Leu Phe
Ala Tyr 1 5 10 33410PRTArtificial
SequenceCDRH3 corresponding to residues 98-107 of SEQ ID NO.157
334Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr 1 5
10 335113PRTArtificial SequenceAmino acid sequence alignement of
SEQ ID NO.16 and SEQ ID NO.178MISC_FEATURE(9)..(9)Xaa is an
amino acid found at a corresponding position in SEQ ID NO.16 or
in SEQ ID NO.178MISC_FEATURE(15)..(15)Xaa is a conservative amino acid
subsitution of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(17)..(17)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(18)..(18)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(19)..(19)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.16 or in SEQ ID
NO.178MISC_FEATURE(21)..(21)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(49)..(49)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.16 or in SEQ ID
NO.178MISC_FEATURE(64)..(64)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(69)..(69)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.16 or in SEQ ID
NO.178MISC_FEATURE(84)..(84)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(89)..(89)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(91)..(91)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.16 or in SEQ ID
NO.178MISC_FEATURE(93)..(93)Xaa is a conservative amino acid subsitution
of a corresponding amino acid in SEQ ID
NO.16MISC_FEATURE(106)..(106)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.16 or in SEQ ID
NO.178MISC_FEATURE(112)..(112)Xaa is a conservative amino acid
subsitution of a corresponding amino acid in SEQ ID NO.16
335Asp Ile Val Met Thr Gln Ser Pro Xaa Ser Leu Ala Val Ser Xaa Gly 1
5 10 15 Xaa Xaa Xaa Thr
Xaa Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 20
25 30 Asn Phe Gln Lys Asn Phe Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln 35 40
45 Xaa Pro Lys Leu Leu Ile Tyr Phe Ala Ser Thr Arg Glu Ser
Ser Xaa 50 55 60
Pro Asp Arg Phe Xaa Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 65
70 75 80 Ile Ser Ser Xaa Gln
Ala Glu Asp Xaa Ala Xaa Tyr Xaa Cys Gln Gln 85
90 95 His Tyr Ser Thr Pro Leu Thr Phe Gly Xaa
Gly Thr Lys Leu Glu Xaa 100 105
110 Lys 336113PRTArtificial SequenceAmino acid sequence
alignement of SEQ ID NO.18 and SEQ ID
NO.179MISC_FEATURE(5)..(5)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.18 or SEQ ID NO.179MISC_FEATURE(8)..(8)Xaa
is an amino acid found at a corresponding position in SEQ ID
NO.18 or SEQ ID NO.179MISC_FEATURE(11)..(11)Xaa is a conservative
substitution of a corresponding amino acid in SEQ ID
NO.18MISC_FEATURE(12)..(12)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.18 or SEQ ID
NO.179MISC_FEATURE(13)..(13)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.18MISC_FEATURE(19)..(19)Xaa is
an amino acid found at a corresponding position in SEQ ID NO.18
or SEQ ID NO.179MISC_FEATURE(20)..(20)Xaa is a conservative substitution
of a corresponding amino acid in SEQ ID
NO.18MISC_FEATURE(38)..(38)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.18MISC_FEATURE(40)..(40)Xaa is
an amino acid found at a corresponding position in SEQ ID NO.18
or SEQ ID NO.179MISC_FEATURE(42)..(42)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.18 or SEQ ID
NO.179MISC_FEATURE(43)..(43)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.18 or SEQ ID
NO.179MISC_FEATURE(48)..(48)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.18MISC_FEATURE(67)..(67)Xaa is
a conservative substitution of a corresponding amino acid in
SEQ ID NO.18MISC_FEATURE(68)..(68)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.18 or SEQ ID
NO.179MISC_FEATURE(70)..(70)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.18MISC_FEATURE(74)..(74)Xaa is
an amino acid found at a corresponding position in SEQ ID NO.18
or SEQ ID NO.179MISC_FEATURE(76)..(76)Xaa is a conservative substitution
of a corresponding amino acid in SEQ ID
NO.18MISC_FEATURE(91)..(91)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.18MISC_FEATURE(108)..(108)Xaa
is an amino acid found at a corresponding position in SEQ ID
NO.18 or SEQ ID NO.179MISC_FEATURE(109)..(109)Xaa is a conservative
substitution of a corresponding amino acid in SEQ ID NO.18
336Glu Val Gln Leu Xaa Gln Ser Xaa Ala Glu Xaa Xaa Xaa Pro Gly Ala 1
5 10 15 Ser Val Xaa Xaa
Ser Cys Lys Ala Ser Gly Tyr Ile Phe Thr Asp Tyr 20
25 30 Glu Ile His Trp Val Xaa Gln Xaa Pro
Xaa Xaa Gly Leu Glu Trp Xaa 35 40
45 Gly Val Ile Asp Pro Glu Thr Gly Asn Thr Ala Phe Asn Gln
Lys Phe 50 55 60
Lys Gly Xaa Xaa Thr Xaa Thr Ala Asp Xaa Ser Xaa Ser Thr Ala Tyr 65
70 75 80 Met Glu Leu Ser Ser
Leu Thr Ser Glu Asp Xaa Ala Val Tyr Tyr Cys 85
90 95 Met Gly Tyr Ser Asp Tyr Trp Gly Gln Gly
Thr Xaa Xaa Thr Val Ser 100 105
110 Ser 337107PRTArtificial SequenceAmino acid sequence
alignement of SEQ ID NO.24 and SEQ ID
NO.182MISC_FEATURE(5)..(5)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.24MISC_FEATURE(11)..(11)Xaa is
a conservative substitution of a corresponding amino acid in
SEQ ID NO.24MISC_FEATURE(12)..(12)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.24 or in SEQ ID
NO.182MISC_FEATURE(15)..(15)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.24MISC_FEATURE(17)..(17)Xaa is
a conservative substitution of a corresponding amino acid in
SEQ ID NO.24MISC_FEATURE(43)..(43)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID
NO.24MISC_FEATURE(69)..(69)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.24 or in SEQ ID
NO.182MISC_FEATURE(72)..(72)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.24MISC_FEATURE(79)..(79)Xaa is
a conservative substitution of a corresponding amino acid in
SEQ ID NO.24MISC_FEATURE(80)..(80)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.24 or in SEQ ID
NO.182MISC_FEATURE(83)..(85)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.24 or in SEQ ID
NO.182MISC_FEATURE(100)..(100)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.24 or in SEQ ID
NO.182MISC_FEATURE(106)..(106)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.24MISC_FEATURE(107)..(107)Xaa
is a conservative substitution of a corresponding amino acid in
SEQ ID NO.24 337Asp Ile Val Met Xaa Gln Ser Pro Ser Ser Xaa Xaa Ala Ser
Xaa Gly 1 5 10 15
Xaa Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Ile His Asn Phe
20 25 30 Leu Asn Trp Phe Gln
Gln Lys Pro Gly Lys Xaa Pro Lys Thr Leu Ile 35
40 45 Phe Arg Ala Asn Arg Leu Val Asp Gly
Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Xaa Asp Tyr Xaa Leu Thr Ile Ser Ser
Leu Xaa Xaa 65 70 75
80 Glu Asp Xaa Xaa Xaa Tyr Ser Cys Leu Gln Tyr Asp Glu Ile Pro Leu
85 90 95 Thr Phe Gly Xaa
Gly Thr Lys Leu Glu Xaa Xaa 100 105
338116PRTArtificial SequenceAmino acid sequence alignement of SEQ ID
NO.26 and SEQ ID NO.183MISC_FEATURE(10)..(10)Xaa is an amino
acid found at a corresponding position in SEQ ID NO.26 or SEQ
ID NO.183MISC_FEATURE(17)..(17)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.26MISC_FEATURE(25)..(25)Xaa
is a conservative substitution of a corresponding amino acid in
SEQ ID NO.26MISC_FEATURE(41)..(41)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.26 or SEQ ID
NO.183MISC_FEATURE(44)..(45)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.26 or SEQ ID
NO.183MISC_FEATURE(49)..(49)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.26MISC_FEATURE(68)..(69)Xaa is
a conservative substitution of a corresponding amino acid in
SEQ ID NO.26MISC_FEATURE(71)..(71)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID
NO.26MISC_FEATURE(80)..(80)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.26 or SEQ ID
NO.183MISC_FEATURE(82)..(82)Xaa is a conservative substitution of a
corresponding amino acid in SEQ ID NO.26MISC_FEATURE(84)..(84)Xaa is
a conservative substitution of a corresponding amino acid in
SEQ ID NO.26MISC_FEATURE(88)..(89)Xaa is an amino acid found at a
corresponding position in SEQ ID NO.26 or SEQ ID
NO.183MISC_FEATURE(93)..(93)Xaa is an amino acid found at a corresponding
position in SEQ ID NO.26 or SEQ ID NO.183 338Glu Val Gln Leu
Gln Glu Ser Gly Pro Xaa Leu Val Lys Pro Ser Gln 1 5
10 15 Xaa Leu Ser Leu Thr Cys Thr Val Xaa
Gly Phe Ser Ile Thr Ser Gly 20 25
30 Tyr Gly Trp His Trp Ile Arg Gln Xaa Pro Gly Xaa Xaa Leu
Glu Trp 35 40 45
Xaa Gly Tyr Ile Asn Tyr Asp Gly His Asn Asp Tyr Asn Pro Ser Leu 50
55 60 Lys Ser Arg Xaa Xaa
Ile Xaa Gln Asp Thr Ser Lys Asn Gln Phe Xaa 65 70
75 80 Leu Xaa Leu Xaa Ser Val Thr Xaa Xaa Asp
Thr Ala Xaa Tyr Tyr Cys 85 90
95 Ala Ser Ser Tyr Asp Gly Leu Phe Ala Tyr Trp Gly Gln Gly Thr
Leu 100 105 110 Val
Thr Val Ser 115
* * * * *
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