U.S. patent application number 14/974675 was filed with the patent office on 2016-07-07 for antigen binding proteins.
The applicant listed for this patent is Glaxo Group Limited. Invention is credited to Paul Algate, Stephanie Jane Clegg, Jennifer L. Craigen, Paul Andrew Hamblin, Alan Peter Lewis, Patrick Mayes, Radha Shah Parmar, Trevor Anthony Kenneth Wattam.
Application Number | 20160193358 14/974675 |
Document ID | / |
Family ID | 49380326 |
Filed Date | 2016-07-07 |
United States Patent
Application |
20160193358 |
Kind Code |
A1 |
Algate; Paul ; et
al. |
July 7, 2016 |
ANTIGEN BINDING PROTEINS
Abstract
The present invention concerns antigen binding proteins and
fragments thereof which specifically bind B Cell Maturation Antigen
(BCMA), particularly human BCMA (hBCMA) and which inhibit the
binding of BAFF and APRIL to the BCMA receptor. Further disclosed
are pharmaceutical compositions, screening and medical treatment
methods.
Inventors: |
Algate; Paul; (Issaquah,
WA) ; Clegg; Stephanie Jane; (Stevenage, GB) ;
Craigen; Jennifer L.; (Stevenage, GB) ; Hamblin; Paul
Andrew; (Stevenage, GB) ; Lewis; Alan Peter;
(Stevenage, GB) ; Mayes; Patrick; (Collegeville,
PA) ; Parmar; Radha Shah; (Stevenage, GB) ;
Wattam; Trevor Anthony Kenneth; (Stevenage, GB) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Glaxo Group Limited |
Brentford |
|
GB |
|
|
Family ID: |
49380326 |
Appl. No.: |
14/974675 |
Filed: |
December 18, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13795314 |
Mar 12, 2013 |
9273141 |
|
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14974675 |
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PCT/EP2012/059762 |
May 24, 2012 |
|
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13795314 |
|
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61490732 |
May 27, 2011 |
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Current U.S.
Class: |
424/183.1 ;
530/391.7; 530/391.9 |
Current CPC
Class: |
C07K 2317/76 20130101;
A61K 47/6849 20170801; A61K 2039/505 20130101; C07K 16/2878
20130101; C07K 2317/92 20130101; C07K 2317/41 20130101; A61P 35/02
20180101; C07K 16/3061 20130101; C07K 2317/24 20130101; C07K
16/2803 20130101; A61K 38/05 20130101; C07K 2317/33 20130101; C07K
2317/73 20130101; C07K 2317/77 20130101; C07K 2317/56 20130101;
A61K 47/6803 20170801; C07K 2317/732 20130101 |
International
Class: |
A61K 47/48 20060101
A61K047/48; A61K 38/05 20060101 A61K038/05; C07K 16/28 20060101
C07K016/28 |
Claims
1. An immunoconjugate comprising an antibody and a cytotoxic agent
wherein said antibody comprises a heavy chain variable region set
forth in SEQ. ID. NO:23 and a light chain variable region set forth
in SEQ. ID. NO:31 and wherein said cytotoxic agent is selected from
monomethylauristatin E (MMAE) and monomethylauristatin F
(MMAF).
2. The immunoconjugate of claim 1 wherein the antibody is an IgG1
isotype.
3. The immunoconjugate of claim 1 wherein the antibody binds BCMA
with an affinity of stronger than 150 pM.
4. The immunoconjugate claim 1 wherein the cytotoxic agent is
covalently bound to said antibody.
5. The immunoconjugate claim 1 wherein the cytotoxic agent is bound
to said antibody via a linker.
6. The immunoconjugate of claim 5 wherein said linker is a
cleavable linker.
7. The immunoconjugate of claim 5 wherein said linker is a
non-cleavable linker.
8. The immunoconjugate of claim 5 wherein the linker is selected
from 6-maleimidocaproyl (MC), maleimidopropanoyl (MP),
valine-citrulline (val-cit), alanine-phenylalanine (ala-phe),
p-aminobenzyloxycarbonyl (PAB), N-Succinimidyl
4-(2-pyridylthio)pentanoate (SPP), N-succinimidyl
4-(N-maleimidomethyl)cyclohexane-1 carboxylate (SMCC), and
N-Succinimidyl (4-iodo-acetyl) aminobenzoate (SIAB).
9. The immunoconjugate of claim 1 wherein said immunoconjugate is
engulfed by a tumor cell when contacted with a tumor cell.
10. The immunoconjugate of claim 1 wherein said antibody is
afucosylated.
11. A pharmaceutical composition comprising the immunoconjugate of
claim 1 and a pharmaceutically acceptable carrier.
12. A method of treating a human patient afflicted with a B cell
lymphoma comprising administering to said human a therapeutically
affective amount of the immunoconjugate of claim 1.
13. The method of claim 12 wherein the B cell lymphoma is selected
from Multiple Myeloma (MM) and Chronic Lymphocytic Leukaemia
(CLL).
14. An immunoconjugate comprising an antibody and a cytoxic agent
wherein said antibody comprises the heavy chain amino acid sequence
set forth in SEQ ID NO:55 and the light chain amino acid sequence
set forth in SEQ ID NO:63 and the cytotoxic agent is
monomethylauristatin E (MMAE), wherein said MMAE is linked to said
antibody via a valine-citrulline (val-cit) linker.
15. An immunoconjugate comprising an antibody and a cytoxic agent
wherein said antibody comprises the heavy chain amino acid sequence
set forth in SEQ ID NO:55 and the light chain amino acid sequence
set forth in SEQ ID NO:63 and the cytotoxic agent is
monomethylauristatin F (MMAF), wherein said MMAF is linked to said
antibody via a 6-maleimidocaproyl (mc) linker.
Description
[0001] This application is a continuation of U.S. application Ser.
No. 13/795,314 filed Mar. 12, 2013, which is a continuation of
International Application No. PCT/EP2012/059762, filed May 24, 2012
which claims priority to and benefit of U.S. Provisional
Application No. 61/490,732 filed on May 27, 2011 which is
incorporated by reference herein in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to antigen binding proteins
and fragments thereof that specifically bind B cell maturation
antigen (BCMA) and in particular human BCMA (hBCMA).
[0003] The present invention also concerns methods of treating
diseases or disorders with said antigen binding fragments,
pharmaceutical compositions comprising said antigen binding
fragments and methods of manufacture. Other embodiments of the
present invention will be apparent from the description below.
BACKGROUND OF THE INVENTION
[0004] BCMA (CD269 or TNFRSF17) is a member of the TNF receptor
superfamily. It is a non-glycosylated integral membrane receptor
for the ligands BAFF and APRIL. BCMA's ligands can also bind
additional receptors: TACI (Transmembrane Activator and Calcium
modulator and cyclophilin ligand Interactor), which binds APRIL and
BAFF; as well as BAFF-R (BAFF Receptor or BR3), which shows
restricted but high affinity for BAFF. Together, these receptors
and their corresponding ligands regulate different aspects of
humoral immunity, B-cell development and homeostasis.
[0005] BCMA's expression is typically restricted to the B-cell
lineage and is reported to increase in terminal B-cell
differentiation. BCMA is expressed by human plasma blasts, plasma
cells from tonsils, spleen and bone marrow, but also by tonsillar
memory B cells and by germinal centre B cells, which have a
TACI-BAFFR low phenotype (Darce et al, 2007). BCMA is virtually
absent on naive and memory B-cells (Novak et al., 2004a and b). The
BCMA antigen is expressed on the cell surface so is accessible to
the antibody, but is also expressed in the golgi. As suggested by
its expression profile, BCMA signalling, typically linked with
B-cell survival and proliferation, is important in the late stages
of B-cell differentiation, as well as the survival of long lived
bone marrow plasma cells (O'Connor et al., 2004) and plasmablasts
(Avery et al., 2003). Furthermore, as BCMA binds APRIL with high
affinity, the BCMA-APRIL signalling axis is suggested to
predominate at the later stages of B-cell differentiation, perhaps
being the most physiologically relevant interaction.
[0006] Multiple Myeloma (MM) is a clonal B-cell malignancy that
occurs in multiple sites within the bone marrow before spreading to
the circulation; either de novo, or as a progression from
monoclonal gammopathy of undetermined significance (MGUS). It is
commonly characterised by increases in paraprotein and osteoclast
activity, as well as hypercalcaemia, cytopenia, renal dysfunction,
hyperviscosity and peripheral neuropathy. Decreases in both normal
antibody levels and numbers of neutrophils are also common, leading
to a life threatening susceptibility to infection. BCMA has been
implicated in the growth and survival of myeloma cell lines in
vitro (Novak et al., 2004a and b; Moreaux et al., 2004).
[0007] BCMA expression (both transcript and protein) is reported to
correlate with disease progression in MM. Using Affymetrix
microarrays, it was demonstrated that the TACI and BCMA genes were
over-expressed in Multiple Myeloma Cells (MMC) compared with their
normal counterparts (Moreaux et al, 2004). Gene expression analysis
has been used to compare human myeloma cells with purified plasma
cells from patients with MGUS and from normal bone marrow as well
as with primary tumour cells from B-cell lineage leukaemias
(Bellucci et al, 2005). The BCMA gene was highly expressed in all
myeloma samples. Although purified plasma cells from patients with
MGUS had lower expression of BCMA, there was no significant
difference when compared with the expression found in normal plasma
cells or myeloma cells. In contrast, BCMA expression was
significantly lower in B-cell Chronic Lymphocytic Leukaemia (CLL),
pre-B Acute Lymphocytic Leukaemia (ALL) and T-cell ALL (T-ALL).
Mouse models that transgenically over-express BAFF or APRIL have a
significant increase in B-cell lymphomas (Batten et al.,
2004--BAFF; Planelles et al., 2004--APRIL). In humans, excess BAFF
and APRIL have been detected in the sera and micro-environments of
patients with a number of B-cell malignancies, as well as other
B-cell disorders.
[0008] All patent and literature references disclosed within the
present specification are expressly and entirely incorporated
herein by reference.
BRIEF DESCRIPTION OF FIGURES
[0009] FIG. 1: FMAT Binding Assay--Figure showing the results of
the FMAT assay for CA8 antibody binding to human and cyno BCMA
expressing HEK293 cells. Human chimeric CA8 binds well to human and
cyno BCMA expressing cells.
[0010] FIG. 2: ELISA Binding Assay--Figure showing the ELISA
results for CA8 antibodies binding to human and cyno BCMA
recombinant proteins. This clearly shows that human chimeric CA8
antibodies bind to human and cyno BCMA proteins equally.
[0011] FIG. 3: BiaCore Binding Assay--Figure showing the binding of
CA8 to BCMA-Fc, TACI-Fc and BAFF-R-Fc proteins in the Biacore
experiment. CA8 chimera antibody does not bind to TACI or BAFF-R
proteins.
[0012] FIG. 4: Cell binding assay--Figure showing binding of murine
S307118G03, S3222110D07, S332121F02 and S332126E04 to H929 multiple
myeloma cells and S3322110D07, S332121F02 and S332126E04 to the
BCMA transfected ARH77 cells as determined by FACS.
[0013] Multiple myeloma cell line H929 or ARH77-hBCMA 10B5 BCMA
expressing transfectant cells were stained with either murine anti
BCMA antibodies (solid histogram) or murine IgG2a isotype control
(open histograms). Cells were analysed by FACS to detect antibody
bound to the cells.
[0014] FIG. 5: Cell binding assay--Figure showing binding of
chimeric CA8 to a panel of multiple myeloma cell lines as
determined by FACS. Binding to H929, OPM-2, JJN-3 and U266 was
tested by flow cytometry and mean fluorescence intensity (MFI)
values measured to determine binding. Synagis was used as an
irrelevant isotype control.
[0015] FIG. 6: Cell binding assay--Figure showing binding curves of
humanised CA8 variants to BCMA transfected ARH77 cells (A) and
multiple myeloma H929 cells (B) as determined by FACS. Humanised
variants J6M0, J6M1, J6M2, J9M0, J9M1 and J9M2 were tested by flow
cytometry and mean fluorescence intensity (MFI) values measured to
determine binding compared to the CA8 chimera.
[0016] FIG. 7: Ligand neutralisation assays--
[0017] (A and B) Figure showing the ability of CA8 and J6M0 to
neutralise binding of recombinant BAFF or APRIL to recombinant BCMA
coated on an ELISA plate. OD values were used to calculate the
antibody mediated inhibition of the maximal signal achieved by the
relevant ligand alone binding to recombinant BCMA. Data is reported
as percentage inhibition of the maximal signal. Antibodies tested
were chimeric CA8 and humanised CA8 version J6M0 in both wild type
and afucosylated (Potelligent) form.
[0018] (A) Neutralisation of BAFF ligand binding;
(B)--Neutralisation APRIL ligand binding.
[0019] (C)--Figure showing the ability of J6M0 BCMA antibody in
inhibition of BAFF or APRIL induced phosphorylation of NFKappaB in
H929 cells. H-929 cells were washed 3 times to remove any sBCMA and
resuspended in serum free medium. J6M0 potelligent antibody was
added to a 96 well plate to give a final well concentrations up to
100 ug/ml along with BAFF or APRIL ligand to give a final well
concentration of 0.6 or 0.2 ug/ml respectively. H-929 cells were
then plated at 7.5.times.104 cells/well in serum free medium. 30
minutes later the cells were lysed and phosphorylated NFkappaB
levels measured using a MSD pNFkappaB assay. MSD reader 502819.
This is data from one independent experiments. Each data point is
the mean/sd of two replicates.
[0020] FIG. 8: ADCC assay--Figure showing ADCC activity of chimeric
CA8 and defucosylated (Fc enhanced) CA8 with target cells
expressing BCMA.
[0021] Human NK cells were incubated with europium labelled ARH77
10B5 BCMA transfected target cells in the presence of varying
concentrations of antibody. Europium release from the target cells
was measured and specific lysis calculated. (A) ADCC dose response
curves of chimeric CA8 compared to isotype control. (B) ADCC dose
response curves for chimeric CA8 and defucosylated chimeric CA8 (Fc
enhanced), against the BCMA expressing cell line ARH77 10B5.
[0022] FIG. 9: ADCC assay--Figure showing ADCC assay on CA8
humanised antibodies using ARH77 BCMA expressing target cells.
[0023] Human PBMC were incubated with europium labelled ARH77 BCMA
transfected target cells in the presence of a range of
concentrations of the J5, J6, J7, J8 or J9 series of humanised CA8
antibodies. Europium release from the target cells was measured and
specific lysis calculated. EC50 values are shown in ug/ml.
[0024] FIG. 10: ADCC assay--Figure showing ADCC activity of
chimeric, S332121F02 (A), S3322110D07 (B) S307118G03 (C) and
humanised S307118G03 H3L0 (D) against ARH7710B5 target cells with
purified NK cells as effector cells. Human NK target cells were
incubated with europium labelled ARH77 10B5 BCMA transfected target
cells in the presence of varying concentrations of antibody.
Europium release from the target cells was measured and specific
lysis calculated.
[0025] FIG. 11: Viability assay dose response curves--Figure
showing dose response curves in a cell viability assay for chimeric
CA8 antibody, chimeric CA8-vcMMAE and chimeric CA8-mcMMAF
antibody-drug conjugates in human multiple myeloma cell lines (A)
NCI-H929 (B) U266-B1 (C) JJN3 and (D) OPM2. Antibody was added to
the cells and the number of viable cells after 96 hours measured
using CelltiterGlo.Data points represent the mean of triplicate
CellTiterGlo measurements. Error bars represent standard error.
[0026] FIG. 12: Impact of CA8 chimeric antibody on cell cycle.
[0027] (A) Cell cycle histograms of NCI-H929 cells treated with
unconjugated chimeric CA8, chimeric CA8-vcMMAE ADC or chimeric
CA8-mcMMAF ADC at 50 ng/mL for the timepoints indicated. Pactitaxel
(100 nM) was used as a positive control for G2/M cell cycle arrest
and cell death. Control human IgG1 was used as a negative control.
Cell cycle analysis was carried out at the times shown on the
graphs. (B) Quantification of the 4N DNA cell population indicative
of G2/M arrest and (C) sub-2N DNA cell population indicative of
cell death for each of the treatments indicated. Cells were seeded
in 12-well plates (2.times.10.sup.5 cells per well in 1 mL of
RPMI+10% FBS). Antibody or ADC was added 6 hours after cell
seeding.
[0028] FIG. 13: Impact of chimeric CA8 on phospho-histone H3.
[0029] Chimeric CA8 ADC treatment results in increased
phospho-Histone H3 staining of NCI-H929 cells. (A,B) Dot plots of
cells stained with propidium iodide to measure DNA content (FL3-H)
x-axis and anti-phospho-Histone H3 (Thr11) antibody (FL1-H) y-axis
after treatment with either Control IgG (A) or chimeric CA8-mcMMAF
(B). (C) Quantification of phospho-Histone H3 positive NCI-H929
cells after a 48 hour treatment with the indicated concentrations
of chimeric CA8 ADCs. Pactitaxel (100 nM) was used as a positive
control for mitotic arrest and control chimera IgG1 was used as a
negative control. Cells were seeded in 12-well plates
(2.times.10.sup.5 cells per well in 1 mL of RPMI+10% FBS). Antibody
or ADC was added 6 hours after cell seeding.
[0030] FIG. 14: Impact of chimeric CA8 on Annexin-V.
[0031] Chimeric CA8 ADC treatment results in increased Annexin-V
staining of NCI-H929 cells.
[0032] (A) Histograms of Annexin-V-FITC (FL1-H; top panels) and
Live cell propidium iodide staining (FL3-H; bottom panels) after
treatment with increasing concentrations of chimeric CA8 ADCs (B)
Quantification of Annexin-V positive NCI-H929 cells after a 96 hour
treatment with the indicated concentrations of chimeric CA8 ADCs.
Pactitaxel (100 nM) was used as a positive control for apoptosis
and control chimera IgG1 was used as a negative control. Cells were
seeded in 12-well plates (2.times.10.sup.5 cells per well in 1 mL
of RPMI+10% FBS). Antibody or ADC was added 6 hours after cell
seeding.
[0033] FIG. 15: Viability assay dose response curves--Figure
showing dose response curves for the unconjugated (Naked) and
vcMMAE and mcMMAF antibody-drug conjugates of chimeric CA8 or
humanized J6M0 antibodies. Antibody drug conjugates were tested
against human multiple myeloma cell lines NCI-H929 and OPM2.
[0034] FIG. 16: Viability assay dose response curves--Figure
showing dose response curves for the unconjugated antibodies,
vcMMAE and mcMMAF antibody-drug conjugates of murine anti-BCMA
antibodies S332121F02, S322110D07, S332126E04 and S307118G03 in
human multiple myeloma cell lines NCI-H929 and U266-B1.
[0035] FIG. 17 ADCC activity of ADC J6M0 molecules--Figure showing
ADCC assay on J6M0 antibodies using ARH77 BCMA expressing target
cells. Human PBMC were incubated with europium labelled ARH77 BCMA
transfected target cells in the presence of a range of
concentrations of J6M0 VVT and potelligent BCMA antibodies
conjugated to MMAE, MMAF, or unconjugated Europium release was
monitored on the Victor 2 1420 multilabel reader.
[0036] FIG. 18 ADCC dose response curves of CA8 J6M0 Potelligent
against a panel of 5 multiple myeloma lines--Human PBMC were
incubated with multiple myeloma target cells in the presence of
varying concentrations of CA8 J6M0 potelligent antibody at an E:T
ratio of 50:1 for 18 hours. The percentage of target cells
remaining in the effecter plus target mixture was then measured by
FACS using a fluorescently labelled anti-CD138 antibody to detect
the target cells and the percent cytotoxicity calculated. A)
Example dose response curves for CA8 J6M0 potelligent against the
five multiple myeloma cell lines tested. Each data point is from a
singlicate value.
[0037] FIG. 19 Effect of dose escalation of J6M0 and drug
conjugated J6M0 on the growth and establishment of NCI-H929 cells
in CB.17 SCID mice Calculated tumour volumes of NCI-H929 tumours in
CB17 SCID mice following twice weekly intraperitoneal dosing of
either 50 or 100 ug J6M0 anti-BCMA or IgG1 isotype control
unconjugated, or conjugated to MMAE or MMAF for 2 weeks. Data
points represent mean tumour volume of n=5 per group
[0038] FIG. 20--Determination of soluble BCMA levels in serum from
healthy volunteers and myeloma patients. Serum samples were
collected from MM patient samples were from a variety of stages
(progressive disease, remission, relapsed, newly diagnosed, and
others). The samples shown in the figure are those from serum
diluted 1/500 prior to the assay.
[0039] A Human BCMA/TNFRSF17 sandwich ELISA kit from R& D
Systems which measures soluble human BCMA levels was used to detect
BCMA following the standard protocol provided with the kit.
SUMMARY OF THE INVENTION
[0040] The present invention provides antigen binding proteins
which bind to membrane bound targets and wherein the antigen
binding protein is capable of internalisation. In a further
embodiment there is provided an immunoconjugate comprising the
antigen binding protein of the present invention and a cytotoxic
agent. In a further embodiment the antigen binding protein has ADCC
effector function for example the antigen binding protein has
enhanced ADCC effector function.
[0041] The present invention provides antigen binding proteins
which specifically bind to BCMA, for example antibodies which
specifically bind to BCMA and which inhibit the binding of BAFF
and/or APRIL to the BCMA receptor. The present invention also
provides antigen binding proteins which specifically bind to BCMA
and which inhibits the binding of BAFF and/or APRIL to BCMA wherein
the antigen binding protein is capable of binding to Fc.gamma.RIIIA
or is capable of Fc.gamma.RIIIA mediated effector function.
[0042] The antigen binding proteins of the present invention
specifically bind to BCMA and inhibit the binding of BAFF and/or
APRIL to BCMA wherein the antigen binding protein has enhanced
binding to Fc.gamma.RIIIA or has enhanced Fc.gamma.RIIIA mediated
effector function. In one embodiment the antigen binding protein is
capable of internalisation.
[0043] In one aspect of the invention there is provided an antigen
binding protein according to the invention as herein described
which binds to non-membrane bound BCMA, for example to serum
BCMA.
[0044] In one embodiment of the present invention there is provided
an immunoconjugate comprising the antigen binding protein of the
present invention and a cytotoxic agent.
[0045] In a further embodiment the antigen binding proteins are
conjugated to a toxin such as an auristatin. In yet a further
embodiment the drug conjugate is vcMMAE or mcMMAF.
[0046] The antigen binding proteins of the present invention are
related to, or derived from a murine monoclonal antibody CA8. The
CA8 murine heavy chain variable region amino acid sequence is
provided as SEQ ID NO. 7 and the CA8 murine light chain variable
region amino acid sequence is provided as SEQ ID NO. 9.
[0047] The antigen binding proteins of the present invention are
related to, or derived from a murine monoclonal antibody
S336105A07. The S336105A07 murine heavy chain variable region amino
acid sequence is provided as SEQ ID NO. 140 and the S336105A07
murine light chain variable region amino acid sequence is provided
as SEQ ID NO. 144.
[0048] Other murine monoclonal antibodies from which antigen
binding proteins of the present invention may also be derived are
included in Table C.
[0049] The heavy chain variable regions (VH) of the present
invention may comprise the following CDRs or variants of these
CDR's (as defined by Kabat (Kabat et al; Sequences of proteins of
Immunological Interest NIH, 1987)):
CDRH1 is provided as SEQ ID NO. 1 or SEQ ID NO. 182 CDRH2 is
provided as SEQ ID NO. 2 or SEQ ID NO. 183 CDRH3 is provided as SEQ
ID NO. 3 or SEQ ID NO. 184
[0050] The light chain variable regions (VL) of the present
invention may comprise the following CDRs or variants of these
CDR's (as defined by Kabat (Kabat et al; Sequences of proteins of
Immunological Interest NIH, 1987)):
CDRL1 is provided as SEQ ID NO. 4 or SEQ ID NO. 185 CDRL2 is
provided as SEQ ID NO. 5 or SEQ ID NO. 186 CDRL3 is provided as SEQ
ID NO. 6 or SEQ ID NO. 187
[0051] The invention also provides a polynucleotide sequence
encoding a heavy chain variable region of any of the
antigen-binding proteins described herein, and a polynucleotide
encoding a light chain variable region of any of the
antigen-binding proteins described herein.
[0052] The invention also provides a polynucleotide sequence
encoding a heavy chain of any of the antigen-binding proteins
described herein, and a polynucleotide encoding a light chain of
any of the antigen-binding proteins described herein.
[0053] Such polynucleotides represent the coding sequence which
corresponds to the equivalent polypeptide sequences, however it
will be understood that such polynucleotide sequences could be
cloned into an expression vector along with a start codon, an
appropriate signal sequence and a stop codon.
[0054] The invention also provides a recombinant transformed or
transfected host cell comprising one or more polynucleotides
encoding a heavy chain and/or a light chain of any of the
antigen-binding proteins described herein.
[0055] The invention further provides a method for the production
of any of the antigen-binding proteins described herein which
method comprises the step of culturing a host cell comprising a
first and second vector, said first vector comprising a
polynucleotide encoding a heavy chain of any of the antigen-binding
proteins described herein and said second vector comprising a
polynucleotide encoding a light chain of any of the antigen-binding
proteins described herein, in a suitable culture media, for example
serum-free culture media.
[0056] The invention further provides a pharmaceutical composition
comprising an antigen-binding protein as described herein and a
pharmaceutically acceptable carrier.
[0057] In a further aspect, the present invention provides a method
of treatment or prophylaxis of a disease or disorder responsive to
inhibiting or blocking BCMA such as the modulation of the
interaction between BCMA and its ligands, BAFF or APRIL which
method comprises the step of administering to said patient a
therapeutically effective amount of the antigen binding protein
thereof as described herein.
[0058] It is therefore an object of the present invention to
provide a therapeutic approach to the treatment of B cell related
disorders or diseases such as antibody mediated or plasma cell
mediated diseases or plasma cell malignancies such as for example
Multiple Myeloma (MM). In particular it is an object of the present
invention to provide antigen binding proteins, especially
antibodies that specifically bind BCMA (e.g. hBCMA) and modulate
(i.e. inhibit or block) the interaction between BCMA and its
ligands such as BAFF and/or APRIL in the treatment of diseases and
disorders responsive to modulation of that interaction.
[0059] In another aspect of the present invention there is provided
a method of treating a human patient afflicted with a B cell
related disorders or diseases such as antibody mediated or plasma
cell mediated diseases or plasma cell malignancies such as for
example Multiple Myeloma (MM) which method comprises the step of
administering to said patient a therapeutically effective amount of
the antigen binding protein as described herein.
[0060] In another aspect of the present invention there is provided
a method of treating a human patient afflicted with Rheumatoid
Arthritis, Psoriasis, Type 1 Diabetes Mellitus or Multiple
Sclerosis which method comprises the step of administering to said
patient a therapeutically effective amount of the antigen binding
protein as described herein.
DETAILED DESCRIPTION OF THE INVENTION
[0061] The present invention provides antigen binding proteins
which bind to membrane bound targets and wherein the antigen
binding protein is capable of internalisation. In a further
embodiment there is provided an immunoconjugate comprising the
antigen binding protein of the present invention and a cytotoxic
agent. In a further embodiment the antigen binding protein has ADCC
effector function for example the antigen binding protein has
enhanced ADCC effector function.
[0062] In one such embodiment there is provided antigen binding
proteins or fragments thereof which specifically bind to BCMA, for
example which specifically binds human BCMA (hBCMA) and which
inhibit the binding of BAFF and/or APRIL to the BCMA receptor.
[0063] In a further embodiment the antigen binding proteins or
fragments of the present invention specifically bind to BCMA and
inhibit the binding of BAFF and/or APRIL to BCMA wherein the
antigen binding proteins or fragments thereof have the ability to
bind to Fc.gamma.RIIIA and mediate FcgRIIIA mediated effector
functions, or have enhanced Fc.gamma.RIIIA mediated effector
function. In one embodiment of the invention as herein provided the
antigen binding proteins are capable of internalisation.
[0064] In one aspect of the invention there is provided an antigen
binding protein according to the invention as herein described
which binds to non-membrane bound BCMA, for example to serum
BCMA.
[0065] In one aspect of the invention there is provided an antigen
binding protein as herein described wherein the antigen binding
protein comprises CDRH3 of SEQ ID NO.3 or a variant of SEQ ID NO.
3.
[0066] In a further aspect of the invention there is provided an
antigen binding protein as herein described wherein the antigen
binding protein further comprises one or more of: CDR H1 of SEQ.
ID. NO: 1, CDRH2: SEQ. ID. NO: 2: CDRL1: SEQ. ID. NO: 4, CDRL2:
SEQ. ID. NO: 5 and/or CDRL3: SEQ. ID. NO: 6 and or variants
thereof.
[0067] In one aspect of the invention there is provided an antigen
binding protein as herein described wherein the antigen binding
protein comprises CDRH3 of SEQ ID NO.184 or a variant of SEQ ID NO.
184.
[0068] In a further aspect of the invention there is provided an
antigen binding protein as herein described wherein the antigen
binding protein further comprises one or more of: CDR H1 of SEQ.
ID. NO: 182, CDRH2: SEQ. ID. NO: 183: CDRL1: SEQ. ID. NO: 185,
CDRL2: SEQ. ID. NO: 186 and/or CDRL3: SEQ. ID. NO: 187 and or
variants thereof.
[0069] In yet a further aspect the antigen binding protein
comprises CDR H3 of SEQ. ID. NO: 3: CDRH2: SEQ. ID. NO: 2: CDR H1
of SEQ. ID. NO:1: CDRL1: SEQ. ID. NO: 4: CDRL2: SEQ. ID. NO: 5 and
CDRL3: SEQ. ID. NO: 6.
[0070] In yet a further aspect the antigen binding protein
comprises CDR H3 of SEQ. ID. NO: 184: CDRH2: SEQ. ID. NO: 183: CDR
H1 of SEQ. ID. NO:182: CDRL1: SEQ. ID. NO: 185: CDRL2: SEQ. ID. NO:
186 and CDRL3: SEQ. ID. NO: 187.
[0071] The antigen binding proteins of the present invention may
comprise heavy chain variable regions and light chain variable
regions of the invention which may be formatted into the structure
of a natural antibody or functional fragment or equivalent thereof.
An antigen binding protein of the invention may therefore comprise
the VH regions of the invention formatted into a full length
antibody, a (Fab')2 fragment, a Fab fragment, or equivalent thereof
(such as scFV, bi- tri- or tetra-bodies, Tandabs etc.), when paired
with an appropriate light chain. The antibody may be an IgG1, IgG2,
IgG3, or IgG4; or IgM; IgA, IgE or IgD or a modified variant
thereof. The constant domain of the antibody heavy chain may be
selected accordingly. The light chain constant domain may be a
kappa or lambda constant domain. Furthermore, the antigen binding
protein may comprise modifications of all classes e.g. IgG dimers,
Fc mutants that no longer bind Fc receptors or mediate C1q binding.
The antigen binding protein may also be a chimeric antibody of the
type described in WO86/01533 which comprises an antigen binding
region and a non-immunoglobulin region.
[0072] The constant region is selected according to any
functionality required e.g. an IgG1 may demonstrate lytic ability
through binding to complement and/or will mediate ADCC (antibody
dependent cell cytotoxicity).
[0073] The antigen binding proteins of the present invention are
derived from the murine antibody having the variable regions as
described in SEQ ID NO:7 and SEQ ID NO:9 or non-murine equivalents
thereof, such as rat, human, chimeric or humanised variants
thereof, for example they are derived from the antibody having the
variable heavy chain sequences as described in SEQ ID NO:11, SEQ ID
NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ
ID NO:23, SEQ ID NO:25, SEQ ID NO:27 and SEQ ID NO:29 and/or the
variable light chain sequences as described in SEQ ID NO:31, SEQ ID
NO:33 and/or SEQ ID NO:35.
[0074] In another embodiment the antigen binding proteins of the
present invention are derived from an antibody having the variable
heavy chain sequences as described in SEQ ID NO:116 or SEQ ID
NO:118 and/or the variable light chain sequences as described in
SEQ ID NO:120, or SEQ ID NO:122.
[0075] In another embodiment the antigen binding proteins of the
present invention are derived from an antibody having the variable
heavy chain sequences as described in SEQ ID NO:140 and/or the
variable light chain sequences as described in SEQ ID NO:144.
[0076] In one aspect of the invention there is provided an antigen
binding protein comprising an isolated heavy chain variable domain
selected from any one of the following: SEQ ID NO:11, SEQ ID NO:13,
SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID
NO:23, SEQ ID NO:25, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:116 or
SEQ ID NO:118.
[0077] In another aspect of the invention there is provided an
antigen binding protein comprising an isolated light chain variable
domain selected from any one of the following: SEQ ID NO:31, SEQ ID
NO:33 or SEQ ID NO:35, SEQ ID NO:120 or SEQ ID NO:122.
[0078] In a further aspect of the invention there is provided an
antigen binding protein comprising an isolated heavy chain variable
domain selected from any one of the following: SEQ ID NO:11, SEQ ID
NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ
ID NO:23, SEQ ID NO:25, SEQ ID NO:27 and SEQ ID NO:29 and an
isolated light chain variable domain selected from any one of the
following: SEQ ID NO:31, SEQ ID NO:33 and/or SEQ ID NO:35.
[0079] In one aspect the antigen binding protein of the present
invention comprises a heavy chain variable region encoded by SEQ.
ID. NO:23 and a light chain variable region encoded by SEQ. ID.
NO:31 In one aspect the antigen binding protein of the present
invention comprises a heavy chain variable region encoded by SEQ.
ID. NO:27 and a light chain variable region encoded by SEQ. ID.
NO:31. In one aspect the antigen binding protein of the present
invention comprises a heavy chain variable region encoded by SEQ.
ID. NO:29 and a light chain variable region encoded by SEQ. ID.
NO:31.
[0080] In one aspect the antigen binding protein of the present
invention comprises a heavy chain variable region encoded by SEQ.
ID. NO:116 and a light chain variable region encoded by SEQ. ID.
NO:120
[0081] In one aspect the antigen binding protein of the present
invention comprises a heavy chain variable region encoded by SEQ.
ID. NO:118 and a light chain variable region encoded by SEQ. ID.
NO:122
[0082] In one aspect there is provided a polynucleotide encoding an
isolated variable heavy chain said polynucleotide comprising SEQ.
ID. NO. 12, or SEQ. ID. NO. 14, or SEQ. ID. NO. 16, or SEQ. ID. NO.
18, or SEQ. ID. NO. 20, or SEQ. ID. NO. 22, or SEQ. ID. NO. 24, or
SEQ. ID. NO. 26, or SEQ. ID. NO. 28, or SEQ. ID. NO. 30 or SEQ. ID.
NO. 117 or SEQ. ID. NO. 119 or SEQ. ID. NO. 141.
[0083] In one aspect there is provided a polynucleotide encoding an
isolated variable light chain said polynucleotide comprising SEQ.
ID. NO. 32, or SEQ. ID. NO. 34, or SEQ. ID. NO. 36 or SEQ. ID. NO.
121 or SEQ. ID. NO.123 or SEQ. ID. NO. 145.
[0084] In a further aspect there is provided a polynucleotide
encoding an isolated variable heavy chain said polynucleotide
comprising SEQ. ID. NO. 24, or SEQ. ID. NO. 28 or SEQ. ID. NO. 30
and a polynucleotide encoding an isolated variable light chain said
polynucleotide comprising SEQ. ID. NO. 32, or SEQ. ID. NO. 34.
[0085] In yet a further aspect there is provided a polynucleotide
encoding an isolated variable heavy chain said polynucleotide
comprising SEQ. ID. NO. 24 and a polynucleotide encoding an
isolated variable light chain said polynucleotide comprising SEQ.
ID. NO.32.
[0086] In yet a further aspect there is provided a polynucleotide
encoding an isolated variable heavy chain said polynucleotide
comprising SEQ. ID. NO. 117 and a polynucleotide encoding an
isolated variable light chain said polynucleotide comprising SEQ.
ID. NO.121.
[0087] In yet a further aspect there is provided a polynucleotide
encoding an isolated variable heavy chain said polynucleotide
comprising SEQ. ID. NO. 119 and a polynucleotide encoding an
isolated variable light chain said polynucleotide comprising SEQ.
ID. NO.123.
[0088] In yet a further aspect there is provided a polynucleotide
encoding an isolated variable heavy chain said polynucleotide
comprising SEQ. ID. NO. 141 and a polynucleotide encoding an
isolated variable light chain said polynucleotide comprising SEQ.
ID. NO.145.
[0089] In a further aspect the antigen binding protein may comprise
any one of the variable heavy chains as described herein in
combination with any one of the light chains as described
herein.
[0090] In one aspect the antigen binding protein is an antibody or
antigen binding fragment thereof comprising one or more CDR's
according to the invention described herein, or one or both of the
heavy or light chain variable domains according to the invention
described herein. In one embodiment the antigen binding protein
binds primate BCMA. In one such embodiment the antigen binding
protein additionally binds non-human primate BCMA, for example
cynomolgus macaque monkey BCMA.
[0091] In another aspect the antigen binding protein is selected
from the group consisting of a dAb, Fab, Fab', F(ab').sub.2, Fv,
diabody, triabody, tetrabody, miniantibody, and a minibody.
[0092] In one aspect of the present invention the antigen binding
protein is a humanised or chimaeric antibody, in a further aspect
the antibody is humanised.
[0093] In one aspect the antibody is a monoclonal antibody.
[0094] In one aspect of the present invention there is provided an
antibody with the heavy chain sequence as set forth in SEQ ID NO:
55 or SEQ ID NO: 59 or SEQ ID NO: 61.
[0095] In one aspect of the present invention there is provided an
antibody with the light chain sequence as set forth in SEQ ID NO:
63 or SEQ ID NO: 65.
[0096] In a further aspect of the invention there is provided an
antibody with the heavy chain sequence of SEQ ID NO: 55 and a light
chain sequence as set forth in SEQ ID NO: 63.
[0097] In one embodiment there is provided an antigen binding
protein which competes with an antigen binding protein of the
invention as herein described. In one such embodiment there is
therefore provided an antigen binding protein which competes with
an antigen binding protein which comprises the heavy chain variable
sequence of SEQ ID NO 23 and the light chain variable region of SEQ
ID NO 31.
[0098] In a further embodiment there is therefore provided an
antigen binding protein which competes with an antigen binding
protein which comprises a heavy chain variable sequence selected
from one of SEQ ID NO 27, SEQ ID NO 29, SEQ ID NO 116, SEQ ID NO
118 and SEQ ID NO 140 and a light chain variable region selected
from one of SEQ ID NO 31, SEQ ID NO 120, SEQ ID NO 122 and SEQ ID
NO 144.
[0099] In another aspect the antigen binding protein binds to human
BCMA with high affinity for example when measured by Biacore the
antigen binding protein binds to human BCMA with an affinity of 20
nM or less or an affinity of 15 nM or less or an affinity of 5 nM
or less or an affinity of 1000 pM or less or an affinity of 500 pM
or less or an affinity of 400 pM or less, or 300 pM or less or for
example about 120 pM. In a further embodiment the antigen binding
protein binds to human BCMA when measured by Biacore of between
about 100 pM and about 500 pM or between about 100 pM and about 400
pM, or between about 100 pM and about 300 pM. In one embodiment of
the present invention the antigen binding protein binds BCMA with
an affinity of less than 150 pm.
[0100] In one such embodiment, this is measured by Biacore, for
example as set out in Example 4.
[0101] In another aspect the antigen binding protein binds to human
BCMA and neutralises the binding of the ligands BAFF and/or APRIL
to the BCMA receptor in a cell neutralisation assay wherein the
antigen binding protein has an IC50 of between about 1 nM and about
500 nM, or between about 1 nM and about 100 nM, or between about 1
nM and about 50 nM, or between about 1 nM and about 25 nM, or
between about 5 nM and about 15 nM. In a further embodiment of the
present invention the antigen binding protein binds BCMA and
neutralises BCMA in a cell neutralisation assay wherein the antigen
binding protein has an IC50 of about 10 nM.
[0102] In one such embodiment, this is measured by a cell
neutralisation assay, for example as set out in Example 4.6.
[0103] The antigen binding proteins, for example antibodies of the
present invention may be produced by transfection of a host cell
with an expression vector comprising the coding sequence for the
antigen binding protein of the invention. An expression vector or
recombinant plasmid is produced by placing these coding sequences
for the antigen binding protein in operative association with
conventional regulatory control sequences capable of controlling
the replication and expression in, and/or secretion from, a host
cell. Regulatory sequences include promoter sequences, e.g., CMV
promoter, and signal sequences which can be derived from other
known antibodies. Similarly, a second expression vector can be
produced having a DNA sequence which encodes a complementary
antigen binding protein light or heavy chain. In certain
embodiments this second expression vector is identical to the first
except insofar as the coding sequences and selectable markers are
concerned, so to ensure as far as possible that each polypeptide
chain is functionally expressed. Alternatively, the heavy and light
chain coding sequences for the antigen binding protein may reside
on a single vector.
[0104] A selected host cell is co-transfected by conventional
techniques with both the first and second vectors (or simply
transfected by a single vector) to create the transfected host cell
of the invention comprising both the recombinant or synthetic light
and heavy chains. The transfected cell is then cultured by
conventional techniques to produce the engineered antigen binding
protein of the invention. The antigen binding protein which
includes the association of both the recombinant heavy chain and/or
light chain is screened from culture by appropriate assay, such as
ELISA or RIA. Similar conventional techniques may be employed to
construct other antigen binding proteins.
[0105] Suitable vectors for the cloning and subcloning steps
employed in the methods and construction of the compositions of
this invention may be selected by one of skill in the art. For
example, the conventional pUC series of cloning vectors may be
used. One vector, pUC19, is commercially available from supply
houses, such as Amersham (Buckinghamshire, United Kingdom) or
Pharmacia (Uppsala, Sweden). Additionally, any vector which is
capable of replicating readily, has an abundance of cloning sites
and selectable genes (e.g., antibiotic resistance), and is easily
manipulated may be used for cloning. Thus, the selection of the
cloning vector is not a limiting factor in this invention.
[0106] The expression vectors may also be characterized by genes
suitable for amplifying expression of the heterologous DNA
sequences, e.g., the mammalian dihydrofolate reductase gene (DHFR).
Other vector sequences include a poly A signal sequence, such as
from bovine growth hormone (BGH) and the betaglobin promoter
sequence (betaglopro). The expression vectors useful herein may be
synthesized by techniques well known to those skilled in this
art.
[0107] The components of such vectors, e.g. replicons, selection
genes, enhancers, promoters, signal sequences and the like, may be
obtained from commercial or natural sources or synthesized by known
procedures for use in directing the expression and/or secretion of
the product of the recombinant DNA in a selected host. Other
appropriate expression vectors of which numerous types are known in
the art for mammalian, bacterial, insect, yeast, and fungal
expression may also be selected for this purpose.
[0108] The present invention also encompasses a cell line
transfected with a recombinant plasmid containing the coding
sequences of the antigen binding proteins of the present invention.
Host cells useful for the cloning and other manipulations of these
cloning vectors are also conventional. However, cells from various
strains of E. Coli may be used for replication of the cloning
vectors and other steps in the construction of antigen binding
proteins of this invention.
[0109] Suitable host cells or cell lines for the expression of the
antigen binding proteins of the invention include mammalian cells
such as NS0, Sp2/0, CHO (e.g. DG44), COS, HEK, a fibroblast cell
(e.g., 3T3), and myeloma cells, for example it may be expressed in
a CHO or a myeloma cell. Human cells may be used, thus enabling the
molecule to be modified with human glycosylation patterns.
Alternatively, other eukaryotic cell lines may be employed. The
selection of suitable mammalian host cells and methods for
transformation, culture, amplification, screening and product
production and purification are known in the art. See, e.g.,
Sambrook et al., cited above.
[0110] Bacterial cells may prove useful as host cells suitable for
the expression of the recombinant Fabs or other embodiments of the
present invention (see, e.g., Pluckthun, A., Immunol. Rev.,
130:151-188 (1992)). However, due to the tendency of proteins
expressed in bacterial cells to be in an unfolded or improperly
folded form or in a non-glycosylated form, any recombinant Fab
produced in a bacterial cell would have to be screened for
retention of antigen binding ability. If the molecule expressed by
the bacterial cell was produced in a properly folded form, that
bacterial cell would be a desirable host, or in alternative
embodiments the molecule may express in the bacterial host and then
be subsequently re-folded. For example, various strains of E. Coli
used for expression are well-known as host cells in the field of
biotechnology. Various strains of B. Subtilis, Streptomyces, other
bacilli and the like may also be employed in this method.
[0111] Where desired, strains of yeast cells known to those skilled
in the art are also available as host cells, as well as insect
cells, e.g. Drosophila and Lepidoptera and viral expression
systems. See, e.g. Miller et al., Genetic Engineering, 8:277-298,
Plenum Press (1986) and references cited therein.
[0112] The general methods by which the vectors may be constructed,
the transfection methods required to produce the host cells of the
invention, and culture methods necessary to produce the antigen
binding protein of the invention from such host cell may all be
conventional techniques. Typically, the culture method of the
present invention is a serum-free culture method, usually by
culturing cells serum-free in suspension. Likewise, once produced,
the antigen binding proteins of the invention may be purified from
the cell culture contents according to standard procedures of the
art, including ammonium 16eroxidi precipitation, affinity columns,
column chromatography, gel electrophoresis and the like. Such
techniques are within the skill of the art and do not limit this
invention. For example, preparations of altered antibodies are
described in WO 99/58679 and WO 96/16990.
[0113] Yet another method of expression of the antigen binding
proteins may utilize expression in a transgenic animal, such as
described in U.S. Pat. No. 4,873,316. This relates to an expression
system using the animals casein promoter which when transgenically
incorporated into a mammal permits the female to produce the
desired recombinant protein in its milk.
[0114] In a further embodiment of the invention there is provided a
method of producing an antibody of the invention which method
comprises the step of culturing a host cell transformed or
transfected with a vector encoding the light and/or heavy chain of
the antibody of the invention and recovering the antibody thereby
produced.
[0115] In accordance with the present invention there is provided a
method of producing an anti-BCMA antibody of the present invention
which binds to and neutralises the activity of human BCMA which
method comprises the steps of;
providing a first vector encoding a heavy chain of the antibody;
providing a second vector encoding a light chain of the antibody;
transforming a mammalian host cell (e.g. CHO) with said first and
second vectors; culturing the host cell of step (c) under
conditions conducive to the secretion of the antibody from said
host cell into said culture media; recovering the secreted antibody
of step (d).
[0116] Once expressed by the desired method, the antibody is then
examined for in vitro activity by use of an appropriate assay.
Presently conventional ELISA assay formats are employed to assess
qualitative and quantitative binding of the antibody to BCMA.
Additionally, other in vitro assays may also be used to verify
neutralizing efficacy prior to subsequent human clinical studies
performed to evaluate the persistence of the antibody in the body
despite the usual clearance mechanisms.
[0117] The dose and duration of treatment relates to the relative
duration of the molecules of the present invention in the human
circulation, and can be adjusted by one of skill in the art
depending upon the condition being treated and the general health
of the patient. It is envisaged that repeated dosing (e.g. once a
week or once every two weeks or once every 3 weeks) over an
extended time period (e.g. four to six months) maybe required to
achieve maximal therapeutic efficacy.
[0118] In one embodiment of the present invention there is provided
a recombinant transformed, transfected or transduced host cell
comprising at least one expression cassette, for example where the
expression cassette comprises a polynucleotide encoding a heavy
chain of an antigen binding protein according to the invention
described herein and further comprises a polynucleotide encoding a
light chain of an antigen binding protein according to the
invention described herein or where there are two expression
cassettes and the 1.sup.st encodes the light chain and the second
encodes the heavy chain. For example in one embodiment the first
expression cassette comprises a polynucleotide encoding a heavy
chain of an antigen binding protein comprising a constant region or
antigen binding fragment thereof which is linked to a constant
region according to the invention described herein and further
comprises a second cassette comprising a polynucleotide encoding a
light chain of an antigen binding protein comprising a constant
region or antigen binding fragment thereof which is linked to a
constant region according to the invention described herein for
example the first expression cassette comprises a polynucleotide
encoding a heavy chain selected from SEQ. ID. NO:56, or SEQ. ID.
NO: 60 or SEQ. ID. NO: 62 and a second expression cassette
comprising a polynucleotide encoding a light chain selected from
SEQ. ID. NO: 64 or SEQ. ID. NO: 66.
[0119] In another embodiment of the invention there is provided a
stably transformed host cell comprising a vector comprising one or
more expression cassettes encoding a heavy chain and/or a light
chain of the antibody comprising a constant region or antigen
binding fragment thereof which is linked to a constant region as
described herein. For example such host cells may comprise a first
vector encoding the light chain and a second vector encoding the
heavy chain, for example the first vector encodes a heavy chain
selected from SEQ. ID. NO: 55, or SEQ. ID. NO: 59 or SEQ. ID. NO:
61 and a second vector encoding a light chain for example the light
chain of SEQ ID NO: 63 or SEQ. ID. NO: 65. In one such example the
first vector encodes a heavy chain selected from SEQ. ID. NO: 55
and a second vector encoding a light chain for example the light
chain of SEQ ID NO: 63.
[0120] In another embodiment of the present invention there is
provided a host cell according to the invention described herein
wherein the cell is eukaryotic, for example where the cell is
mammalian. Examples of such cell lines include CHO or NS0.
[0121] In another embodiment of the present invention there is
provided a method for the production of an antibody comprising a
constant region or antigen binding fragment thereof which is linked
to a constant region according to the invention described herein
which method comprises the step of culturing a host cell in a
culture media, for example serum-free culture media.
[0122] In another embodiment of the present invention there is
provided a method according to the invention described herein
wherein said antibody is further purified to at least 95% or
greater (e.g. 98% or greater) with respect to said antibody
containing serum-free culture media.
[0123] In yet another embodiment there is provided a pharmaceutical
composition comprising an antigen binding protein and a
pharmaceutically acceptable carrier.
[0124] In another embodiment of the present invention there is
provided a kit-of-parts comprising the composition according to the
invention described herein described together with instructions for
use.
[0125] The mode of administration of the therapeutic agent of the
invention may be any suitable route which delivers the agent to the
host. The antigen binding proteins, and pharmaceutical compositions
of the invention are particularly useful for parenteral
administration, i.e., subcutaneously (s.c.), intrathecally,
intraperitoneally, intramuscularly (i.m.) or intravenously (i.v.).
In one such embodiment the antigen binding proteins of the present
invention are administered intravenously or subcutaneously.
[0126] Therapeutic agents of the invention may be prepared as
pharmaceutical compositions containing an effective amount of the
antigen binding protein of the invention as an active ingredient in
a pharmaceutically acceptable carrier. In one embodiment the
prophylactic agent of the invention is an aqueous suspension or
solution containing the antigen binding protein in a form ready for
injection. In one embodiment the suspension or solution is buffered
at physiological pH. In one embodiment the compositions for
parenteral administration will comprise a solution of the antigen
binding protein of the invention or a cocktail thereof dissolved in
a pharmaceutically acceptable carrier. In one embodiment the
carrier is an aqueous carrier. A variety of aqueous carriers may be
employed, e.g., 0.9% saline, 0.3% glycine, and the like. These
solutions may be made sterile and generally free of particulate
matter. These solutions may be sterilized by conventional, well
known sterilization techniques (e.g., filtration). The compositions
may contain pharmaceutically acceptable auxiliary substances as
required to approximate physiological conditions such as pH
adjusting and buffering agents, etc. The concentration of the
antigen binding protein of the invention in such pharmaceutical
formulation can vary widely, i.e., from less than about 0.5%,
usually at or at least about 1% to as much as about 15 or 20% by
weight and will be selected primarily based on fluid volumes,
viscosities, etc., according to the particular mode of
administration selected.
[0127] Thus, a pharmaceutical composition of the invention for
intravenous infusion could be made up to contain about 250 ml of
sterile Ringer's solution, and about 1 to about 30 or 5 mg to about
25 mg of an antigen binding protein of the invention per ml of
Ringer's solution. Actual methods for preparing parenterally
administrable compositions are well known or will be apparent to
those skilled in the art and are described in more detail in, for
example, Remington's Pharmaceutical Science, 15.sup.th ed., Mack
Publishing Company, Easton, Pa. For the preparation of
intravenously administrable antigen binding protein formulations of
the invention see Lasmar U and Parkins D "The formulation of
Biopharmaceutical products", Pharma. Sci. Tech. today, page
129-137, Vol. 3 (3 Apr. 2000); Wang, W "Instability, stabilisation
and formulation of liquid protein pharmaceuticals", Int. J. Pharm
185 (1999) 129-188; Stability of Protein Pharmaceuticals Part A and
B ed Ahern T. J., Manning M. C., New York, N.Y.: Plenum Press
(1992); Akers, M. J. "Excipient-Drug interactions in Parenteral
Formulations", J. Pharm Sci 91 (2002) 2283-2300; Imamura, K et al
"Effects of types of sugar on stabilization of Protein in the dried
state", J Pharm Sci 92 (2003) 266-274; Izutsu, Kkojima, S.
"Excipient crystallinity and its protein-structure-stabilizing
effect during freeze-drying", J Pharm. Pharmacol, 54 (2002)
1033-1039; Johnson, R, "Mannitol-sucrose mixtures-versatile
formulations for protein peroxidise19g19n", J. Pharm. Sci, 91
(2002) 914-922; and Ha, E Wang W, Wang Y. j. "Peroxide formation in
polysorbate 80 and protein stability", J. Pharm Sci, 91,
2252-2264(2002) the entire contents of which are incorporated
herein by reference and to which the reader is specifically
referred.
[0128] In one embodiment the therapeutic agent of the invention,
when in a pharmaceutical preparation, is present in unit dose
forms. The appropriate therapeutically effective dose will be
determined readily by those of skill in the art. Suitable doses may
be calculated for patients according to their weight, for example
suitable doses may be in the range of about 0.1 to about 20 mg/kg,
for example about 1 to about 20 mg/kg, for example about 10 to
about 20 mg/kg or for example about 1 to about 15 mg/kg, for
example about 10 to about 15 mg/kg. To effectively treat conditions
such as Multiple myeloma, SLE or IPT in a human, suitable doses may
be within the range of about 0.1 to about 1000 mg, for example
about 0.1 to about 500 mg, for example about 500 mg, for example
about 0.1 to about 100 mg, or about 0.1 to about 80 mg, or about
0.1 to about 60 mg, or about 0.1 to about 40 mg, or for example
about 1 to about 100 mg, or about 1 to about 50 mg, of an antigen
binding protein of this invention, which may be administered
parenterally, for example subcutaneously, intravenously or
intramuscularly. Such dose may, if necessary, be repeated at
appropriate time intervals selected as appropriate by a
physician.
[0129] The antigen binding proteins described herein can be
lyophilized for storage and reconstituted in a suitable carrier
prior to use. This technique has been shown to be effective with
conventional immunoglobulins and art-known peroxidise and
reconstitution techniques can be employed.
[0130] In another aspect of the invention there is provided an
antigen binding protein as herein described for use in a
medicament.
[0131] In one aspect of the present invention there is provided an
antigen binding protein according to the invention as herein
described for use in the treatment of rheumatoid arthritis, Type 1
Diabetes Mellitus, multiple sclerosis or psoriasis wherein said
method comprises the step of administering to said patient a
therapeutically effective amount of the antigen binding protein as
described herein.
[0132] In one embodiment of the present invention, methods are
provided for treating cancer in a human comprising administering to
said human an antigen binding protein that specifically binds to
BCMA. In some instances the antigen binding protein is part of an
immunoconjugate.
[0133] In another aspect of the present invention there is provided
an antigen binding protein according to the invention as herein
described for use in the treatment of a B-cell mediated or plasma
cell mediated disease or antibody mediated disease or disorder
selected from Multiple Myeloma (MM), chronic lymphocytic leukemia
(CLL), Non-secretory multiple myeloma, Smoldering multiple myeloma,
Monoclonal gammopathy of undetermined significance (MGUS), Solitary
plasmacytoma (Bone, Extramedullary), Lymphoplasmacytic lymphoma
(LPL), Waldenstrom's Macroglobulinemia, Plasma cell leukemia,
Primary Amyloidosis (AL), Heavy chain disease, Systemic lupus
erythematosus (SLE), POEMS syndrome/osteosclerotic myeloma, Type I
and II cryoglobulinemia, Light chain deposition disease,
Goodpasture's syndrome, Idiopathic thrombocytopenic purpura (ITP),
Acute glomerulonephritis, Pemphigus and Pemphigoid disorders, and
Epidermolysis bullosa acquisita; or any Non-Hodgkin's Lymphoma
B-cell leukemia or Hodgkin's lymphoma (HL) with BCMA expression or
any diseases in which patients develop neutralising antibodies to
recombinant protein replacement therapy wherein said method
comprises the step of administering to said patient a
therapeutically effective amount of the antigen binding protein as
described herein.
[0134] B-cell disorders can be divided into defects of B-cell
development/immunoglobulin production (immunodeficiencies) and
excessive/uncontrolled proliferation (lymphomas, leukemias). As
used herein, B-cell disorder refers to both types of diseases, and
methods are provided for treating B-cell disorders with an antigen
binding protein.
[0135] In a particular aspect, the disease or disorder is selected
from the group consisting of Multiple Myeloma (MM), Chronic
Lymphocytic Leukaemia (CLL), Solitary Plasmacytoma (Bone,
Extramedullary), Waldenstrom's Macroglobulinemia.
[0136] In one aspect of the present invention the disease is
Multiple Myeloma, Smoldering Multiple Myeloma (SMM) or Solitary
Plasmacytoma (Bone, Extramedullary).
[0137] In one aspect of the present invention the disease is
Multiple Myeloma.
[0138] In one aspect of the present invention the disease is
Systemic lupus erythematosus (SLE)
[0139] In one aspect of the present invention the disease is
Idiopathic thrombocytopenic purpura (ITP)
[0140] Use of the antigen binding protein as described herein in
the manufacture of a medicament for the treatment of diseases and
disorders as described herein is also provided.
[0141] For example in one aspect of the invention there is provided
the use of the antigen binding protein as described herein for use
in the treatment or prophylaxis of diseases and disorders
responsive to modulation (such as inhibiting or blocking) of the
interaction between BCMA and the ligands BAFF and APRIL.
[0142] In another aspect of the invention there is provided the use
of the antigen binding protein as described herein for use in the
treatment or prophylaxis of an antibody mediated or plasma cell
mediated disease or disorder selected from rheumatoid arthritis,
Type 1 Diabetes Mellitus, multiple sclerosis or psoriasis.
[0143] In another aspect of the invention there is provided the use
of the antigen binding protein as described herein for use in the
treatment or prophylaxis of an antibody mediated or plasma cell
mediated disease or disorder selected from Multiple Myeloma (MM),
chronic lymphocytic leukemia (CLL), Monoclonal gammopathy of
undetermined significance (MGUS), Smoldering multiple myeloma
(SMM), Solitary Plasmacytoma (Bone, Extramedullary), Waldenstrom's
Macroglobulinemia, Primary Amyloidosis (AL), Heavy chain disease,
Systemic lupus erythematosus (SLE), POEMS syndrome/osteosclerotic
myeloma, Type I and II cryoglobulinemia, Light chain deposition
disease, Goodpastures syndrome, Idiopathic thrombocytopenic purpura
(ITP), Acute glomerulonephritis, Pemphigus and Pemphigoid disorders
and Epidermolysis bullosa acquisita, any Non-Hodgkin Lymphoma and
Leukemia with BCMA expression or any diseases in which patients
develop neutralising antibodies to recombinant protein replacement
therapy wherein said method comprises the step of administering to
said patient a therapeutically effective amount of the antigen
binding protein as described herein.
[0144] In one aspect, the invention provides a pharmaceutical
composition comprising an antigen binding protein of the present
invention or a functional fragment thereof and a pharmaceutically
acceptable carrier for treatment or prophylaxis of rheumatoid
arthritis, Type 1 Diabetes Mellitus, multiple sclerosis or
psoriasis or an antibody mediated or plasma cell mediated disease
or disorder selected from selected from Multiple Myeloma (MM),
chronic lymphocytic leukemia (CLL), Monoclonal gammopathy of
undetermined significance (MGUS), Smoldering multiple myeloma
(SMM), Solitary Plasmacytoma (Bone, Extramedullary), Waldenstrom's
Macroglobulinemia, Primary Amyloidosis (AL), Heavy chain disease,
Systemic lupus erythematosus (SLE), POEMS syndrome/osteosclerotic
myeloma, Type I and II cryoglobulinemia, Light chain deposition
disease, Goodpastures syndrome, Idiopathic thrombocytopenic purpura
(ITP), Acute glomerulonephritis, Pemphigus and Pemphigoid disorders
and Epidermolysis bullosa acquisita, any Non-Hodgkin Lymphoma and
Leukemia with BCMA expression or any diseases in which patients
develop neutralising antibodies to recombinant protein replacement
therapy wherein said method comprises the step of administering to
said patient a therapeutically effective amount of the antigen
binding protein as described herein.
[0145] In another embodiment of the present invention there is
provided a method of treating a human patient afflicted with
rheumatoid arthritis, Type 1 Diabetes Mellitus, multiple sclerosis
or psoriasis or an antibody mediated or plasma cell mediated
disorder or disease which method comprises the step of
administering a therapeutically effective amount of the antigen
binding protein according to the invention as described herein, for
example there is provided a method of treating a human patient
afflicted with an antibody mediated or plasma cell mediated disease
or disorder selected from In another aspect of the present
invention there is provided an antigen binding protein according to
the invention as herein described for use in the treatment of an
antibody mediated or plasma cell mediated disease or disorder
selected from Multiple Myeloma (MM), Chronic Lymphocytic Leukaemia
(CLL) Monoclonal gammopathy of undetermined significance (MGUS),
Smoldering multiple myeloma (SMM), Solitary Plasmacytoma (Bone,
Extramedullary), Waldenstrom's Macroglobulinemia, Primary
Amyloidosis (AL), Heavy chain disease, Systemic lupus erythematosus
(SLE), POEMS syndrome/osteosclerotic myeloma, Type I and II
cryoglobulinemia, Light chain deposition disease, Goodpastures
syndrome, Idiopathic thrombocytopenic purpura (ITP), Acute
glomerulonephritis, Pemphigus and Pemphigoid disorders and
Epidermolysis bullosa acquisita, any Non-Hodgkin Lymphoma and
Leukemia with BCMA expression or any diseases in which patients
develop neutralising antibodies to recombinant protein replacement
therapy wherein said method comprises the step of administering a
pharmaceutical composition comprising an antigen binding protein
according to the invention herein in combination with a
pharmaceutically acceptable carrier.
[0146] In a further embodiment there is provided a method of
treating a human patient afflicted with Multiple Myeloma (MM).
DEFINITIONS
[0147] As used herein, the terms "cancer," "neoplasm," and "tumor"
are used interchangeably and, in either the singular or plural
form, refer to cells that have undergone a malignant transformation
that makes them pathological to the host organism. Primary cancer
cells can be readily distinguished from non-cancerous cells by
well-established techniques, particularly histological examination.
The definition of a cancer cell, as used herein, includes not only
a primary cancer cell, but any cell derived from a cancer cell
ancestor. This includes metastasized cancer cells, and in vitro
cultures and cell lines derived from cancer cells. When referring
to a type of cancer that normally manifests as a solid tumor, a
"clinically detectable" tumor is one that is detectable on the
basis of tumor mass; e.g., by procedures such as computed
tomography (CT) scan, magnetic resonance imaging (MRI), X-ray,
ultrasound or palpation on physical examination, and/or which is
detectable because of the expression of one or more cancer-specific
antigens in a sample obtainable from a patient. Tumors may be a
hematopoietic (or hematologic or hematological or blood-related)
cancer, for example, cancers derived from blood cells or immune
cells, which may be referred to as "liquid tumors." Specific
examples of clinical conditions based on hematologic tumors include
leukemias such as chronic myelocytic leukemia, acute myelocytic
leukemia, chronic lymphocytic leukemia and acute lymphocytic
leukemia; plasma cell malignancies such as multiple myeloma, MGUS
and Waldenstrom's macroglobulinemia; lymphomas such as
non-Hodgkin's lymphoma, Hodgkin's lymphoma; and the like.
[0148] The cancer may be any cancer in which an abnormal number of
blast cells or unwanted cell proliferation is present or that is
diagnosed as a hematological cancer, including both lymphoid and
myeloid malignancies. Myeloid malignancies include, but are not
limited to, acute myeloid (or myelocytic or myelogenous or
myeloblastic) leukemia (undifferentiated or differentiated), acute
promyeloid (or promyelocytic or promyelogenous or promyeloblastic)
leukemia, acute myelomonocytic (or myelomonoblastic) leukemia,
acute monocytic (or monoblastic) leukemia, erythroleukemia and
megakaryocytic (or megakaryoblastic) leukemia. These leukemias may
be referred together as acute myeloid (or myelocytic or
myelogenous) leukemia (AML). Myeloid malignancies also include
myeloproliferative disorders (MPD) which include, but are not
limited to, chronic myelogenous (or myeloid) leukemia (CML),
chronic myelomonocytic leukemia (CMML), essential thrombocythemia
(or thrombocytosis), and polcythemia vera (PCV). Myeloid
malignancies also include myelodysplasia (or myelodysplastic
syndrome or MDS), which may be referred to as refractory anemia
(RA), refractory anemia with excess blasts (RAEB), and refractory
anemia with excess blasts in transformation (RAEBT); as well as
myelofibrosis (MFS) with or without agnogenic myeloid
metaplasia.
[0149] Hematopoietic cancers also include lymphoid malignancies,
which may affect the lymph nodes, spleens, bone marrow, peripheral
blood, and/or extranodal sites. Lymphoid cancers include B-cell
malignancies, which include, but are not limited to, B-cell
non-Hodgkin's lymphomas (B-NHLs). B-NHLs may be indolent (or
low-grade), intermediate-grade (or aggressive) or high-grade (very
aggressive). Indolent Bcell lymphomas include follicular lymphoma
(FL); small lymphocytic lymphoma (SLL); marginal zone lymphoma
(MZL) including nodal MZL, extranodal MZL, splenic MZL and splenic
MZL with villous lymphocytes; lymphoplasmacytic lymphoma (LPL); and
mucosa-associated-lymphoid tissue (MALT or extranodal marginal
zone) lymphoma. Intermediate-grade B-NHLs include mantle cell
lymphoma (MCL) with or without leukemic involvement, diffuse large
cell lymphoma (DLBCL), follicular large cell (or grade 3 or grade
3B) lymphoma, and primary mediastinal lymphoma (PML). High-grade
B-NHLs include Burkitt's lymphoma (BL), Burkitt-like lymphoma,
small non-cleaved cell lymphoma (SNCCL) and lymphoblastic lymphoma.
Other B-NHLs include immunoblastic lymphoma (or immunocytoma),
primary effusion lymphoma, HIV associated (or AIDS related)
lymphomas, and post-transplant lymphoproliferative disorder (PTLD)
or lymphoma. B-cell malignancies also include, but are not limited
to, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia
(PLL), Waldenstrom's macroglobulinemia (WM), hairy cell leukemia
(HCL), large granular lymphocyte (LGL) leukemia, acute lymphoid (or
lymphocytic or lymphoblastic) leukemia, and Castleman's disease.
NHL may also include T-cell non-Hodgkin's lymphoma s(T-NHLs), which
include, but are not limited to T-cell non-Hodgkin's lymphoma not
otherwise specified (NOS), peripheral T-cell lymphoma (PTCL),
anaplastic large cell lymphoma (ALCL), angioimmunoblastic lymphoid
disorder (AILD), nasal natural killer (NK) cell/T-cell lymphoma,
gamma/delta lymphoma, cutaneous T cell lymphoma, mycosis fungoides,
and Sezary syndrome.
[0150] Hematopoietic cancers also include Hodgkin's lymphoma (or
disease) including classical Hodgkin's lymphoma, nodular sclerosing
Hodgkin's lymphoma, mixed cellularity Hodgkin's lymphoma,
lymphocyte predominant (LP) Hodgkin's lymphoma, nodular LP
Hodgkin's lymphoma, and lymphocyte depleted Hodgkin's lymphoma.
Hematopoietic cancers also include plasma cell diseases or cancers
such as multiple myeloma (MM) including smoldering MM, monoclonal
gammopathy of undetermined (or unknown or unclear) significance
(MGUS), plasmacytoma (bone, extramedullary), lymphoplasmacytic
lymphoma (LPL), Waldenstrom's Macroglobulinemia, plasma cell
leukemia, and primary amyloidosis (AL). Hematopoietic cancers may
also include other cancers of additional hematopoietic cells,
including polymorphonuclear leukocytes (or neutrophils), basophils,
eosinophils, dendritic cells, platelets, erythrocytes and natural
killer cells. Tissues which include hematopoietic cells referred
herein to as "hematopoietic cell tissues" include bone marrow;
peripheral blood; thymus; and peripheral lymphoid tissues, such as
spleen, lymph nodes, lymphoid tissues associated with mucosa (such
as the gut-associated lymphoid tissues), tonsils, Peyer's patches
and appendix, and lymphoid tissues associated with other mucosa,
for example, the bronchial linings.
[0151] The term "antigen binding protein" as used herein refers to
antibodies, antibody fragments and other protein constructs which
are capable of binding to and neutralising human BCMA.
[0152] The terms Fv, Fc, Fd, Fab, or F(ab)2 are used with their
standard meanings (see, e.g., Harlow et al., Antibodies A
Laboratory Manual, Cold Spring Harbor Laboratory, (1988)).
[0153] The term "antibody" is used herein in the broadest sense and
specifically covers monoclonal antibodies (including full length
monoclonal antibodies), polyclonal antibodies, multispecific
antibodies (e.g. bispecific antibodies)
[0154] The term "monoclonal antibody" as used herein refers to an
antibody obtained from a population of substantially homogenous
antibodies i.e. the individual antibodies comprising the population
are identical except for possible naturally occurring mutations
that may be present in minor amounts. Monoclonal antibodies are
highly specific being directed against a single antigenic binding
site. Furthermore, in contrast to polyclonal antibody preparations
which typically include different antibodies directed against
different determinants (epitopes), each monoclonal antibody is
directed against a single determinant on the antigen.
[0155] A "chimeric antibody" refers to a type of engineered
antibody in which a portion of the heavy and/or light chain is
identical with or homologous to corresponding sequences in
antibodies derived from a particular donor antibody class or
subclass, while the remainder of the chain(s) is identical with or
homologous to corresponding sequences in antibodies derived from
another species or belonging to another antibody class or subclass,
as well as fragments of such antibodies, so long as they exhibit
the desired biological activity (U.S. Pat. No. 4,816,567 and
Morrison et al. Proc. Natl. Acad. Sci. USA 81:6851-6855)
(1984)).
[0156] A "humanised antibody" refers to a type of engineered
antibody having its CDRs derived from a non-human donor
immunoglobulin, the remaining immunoglobulin-derived parts of the
molecule being derived from one (or more) human immunoglobulin(s).
In addition, framework support residues may be altered to preserve
binding affinity (see, e.g., Queen et al., Proc. Natl Acad Sci USA,
86:10029-10032 (1989), Hodgson et al., Bio/Technology, 9:421
(1991)). A suitable human acceptor antibody may be one selected
from a conventional database, e.g., the KABAT.RTM. database, Los
Alamos database, and Swiss Protein database, by homology to the
nucleotide and amino acid sequences of the donor antibody. A human
antibody characterized by a homology to the framework regions of
the donor antibody (on an amino acid basis) may be suitable to
provide a heavy chain constant region and/or a heavy chain variable
framework region for insertion of the donor CDRs. A suitable
acceptor antibody capable of donating light chain constant or
variable framework regions may be selected in a similar manner. It
should be noted that the acceptor antibody heavy and light chains
are not required to originate from the same acceptor antibody. The
prior art describes several ways of producing such humanised
antibodies--see for example EP-A-0239400 and EP-A-054951.
[0157] For nucleic acids, the term "substantial identity" indicates
that two nucleic acids, or designated sequences thereof, when
optimally aligned and compared, are identical, with appropriate
nucleotide insertions or deletions, in at least about 80% of the
nucleotides, at least about 90% to about 95%, or at least about 98%
to about 99.5% of the nucleotides. Alternatively, substantial
identity exists when the segments will hybridize under selective
hybridization conditions, to the complement of the strand.
"Identity," means, for polynucleotides and polypeptides, as the
case may be, the comparison calculated using an algorithm provided
in (1) and (2) below: [0158] (1) Identity for polynucleotides is
calculated by multiplying the total number of nucleotides in a
given sequence by the integer defining the percent identity divided
by 100 and then subtracting that product from said total number of
nucleotides in said sequence, or:
[0158] nn.ltoreq.xn-(xny),
wherein nn is the number of nucleotide alterations, xn is the total
number of nucleotides in a given sequence, y is 0.95 for 95%, 0.97
for 97% or 1.00 for 100%, and is the symbol for the multiplication
operator, and wherein any non-integer product of xn and y is
rounded down to the nearest integer prior to subtracting it from
xn. Alterations of a polynucleotide sequence encoding a polypeptide
may create nonsense, missense or frameshift mutations in this
coding sequence and thereby alter the polypeptide encoded by the
polynucleotide following such alterations. [0159] (2) Identity for
polypeptides is calculated by multiplying the total number of amino
acids by the integer defining the percent identity divided by 100
and then subtracting that product from said total number of amino
acids, or:
[0159] na.ltoreq.xa-(xay),
wherein na is the number of amino acid alterations, xa is the total
number of amino acids in the sequence, y is 0.95 for 95%, 0.97 for
97% or 1.00 for 100%, and is the symbol for the multiplication
operator, and wherein any non-integer product of xa and y is
rounded down to the nearest integer prior to subtracting it from
xa
[0160] For nucleotide and amino acid sequences, the term
"identical" indicates the degree of identity between two nucleic
acid or amino acid sequences when optimally aligned and compared
with appropriate insertions or deletions.
[0161] "Isolated" means altered "by the hand of man" from its
natural state, has been changed or removed from its original
environment, or both. For example, a polynucleotide or a
polypeptide naturally present in a living organism is not
"isolated," but the same polynucleotide or polypeptide separated
from the coexisting materials of its natural state is "isolated",
including but not limited to when such polynucleotide or
polypeptide is introduced back into a cell, even if the cell is of
the same species or type as that from which the polynucleotide or
polypeptide was separated.
[0162] Throughout the present specification and the accompanying
claims the term "comprising" and "comprises" incorporates
"consisting of" and "consists of". That is, these words are
intended to convey the possible inclusion of other elements or
integers not specifically recited, where the context allows.
[0163] The term "specifically binds" as used throughout the present
specification in relation to antigen binding proteins of the
invention means that the antigen binding protein binds human BCMA
(hBCMA) with no or insignificant binding to other human proteins.
The term however does not exclude the fact that antigen binding
proteins of the invention may also be cross-reactive with other
forms of BCMA, for example primate BCMA. For example in one
embodiment the antigen binding protein does not bind to TACI or
BAFF-R.
[0164] The term "inhibits" as used throughout the present
specification in relation to antigen binding proteins of the
invention means that the biological activity of BCMA is reduced in
the presence of the antigen binding proteins of the present
invention in comparison to the activity of BCMA in the absence of
such antigen binding proteins. Inhibition may be due but not
limited to one or more of blocking ligand binding, preventing the
ligand activating the receptor, and/or down regulating the BCMA.
Inhibits can also refer to an antigen binding protein binding to
BCMA and causing cell apoptosis or ADCC. The antibodies of the
invention may neutralise the activity of the BCMA ligands BAFF
and/or APRIL binding to BCMA. Levels of neutralisation can be
measured in several ways, for example by use of the assays as set
out in the examples below, for example in 4.4 in an H929 cell NFkB
signalling assay. The BCMA ligands BAFF and APRIL are able to
induce NFkB signalling and downstream events following binding to
BCMA. The neutralisation of BCMA in this assay is measured by
assessing the ability of anti-BCMA monoclonal antibodies to inhibit
BAFF or APRIL driven NFkB induction.
[0165] If an antibody or antigen binding fragment thereof is
capable of neutralisation then this is indicative of inhibition of
the interaction between human BAFF or APRIL and BCMA. Antibodies
which are considered to have neutralising activity against human
BCMA would have an IC50 of less than 30 micrograms/ml, or less than
20 micrograms/ml, or less than 10 micrograms/ml, or less than 5
micrograms/ml or less than 1 micrograms/ml or less than 0.1
micrograms/ml in the H929 stimulation assay as set out in Example
4.4
[0166] "CDRs" are defined as the complementarity determining region
amino acid sequences of an antibody which are the hypervariable
domains of immunoglobulin heavy and light chains. There are three
heavy chain and three light chain CDRs (or CDR regions) in the
variable portion of an immunoglobulin. Thus, "CDRs" as used herein
may refer to all three heavy chain CDRs, or all three light chain
CDRs (or both all heavy and all light chain CDRs, if
appropriate).
[0167] CDRs provide the majority of contact residues for the
binding of the antibody to the antigen or epitope. CDRs of interest
in this invention are derived from donor antibody variable heavy
and light chain sequences, and include analogs of the naturally
occurring CDRs, which analogs also share or retain the same antigen
binding specificity and/or neutralizing ability as the donor
antibody from which they were derived.
[0168] The CDR sequences of antibodies can be determined by the
Kabat numbering system (Kabat et al; (Sequences of proteins of
Immunological Interest NIH, 1987), alternatively they can be
determined using the Chothia numbering system (Al-Lazikani et al.,
(1997) JMB 273, 927-948), the contact definition method (MacCallum
R. M., and Martin A. C. R. and Thornton J. M, (1996), Journal of
Molecular Biology, 262 (5), 732-745) or any other established
method for numbering the residues in an antibody and determining
CDRs known to the skilled man in the art
[0169] Other numbering conventions for CDR sequences available to a
skilled person include "AbM" (University of Bath) and "contact"
(University College London) methods. The minimum overlapping region
using at least two of the Kabat, Chothia, AbM and contact methods
can be determined to provide the "minimum binding unit". The
minimum binding unit may be a sub-portion of a CDR.
[0170] Table A below represents one definition using each numbering
convention for each CDR or binding unit. The Kabat numbering scheme
is used in Table X to number the variable domain amino acid
sequence. It should be noted that some of the CDR definitions may
vary depending on the individual publication used.
TABLE-US-00001 TABLE A Minimum Chothia Contact binding Kabat CDR
CDR AbM CDR CDR unit H1 31-35/35A/35B 26-32/ 26-35/35A/35B
30-35/35A/ 31-32 33/34 35B H2 50-65 52-56 50-58 47-58 52-56 H3
95-102 95-102 95-102 93-101 95-101 L1 24-34 24-34 24-34 30-36 30-34
L2 50-56 50-56 50-56 46-55 50-55 L3 89-97 89-97 89-97 89-96
89-96
[0171] Throughout this specification, amino acid residues in
antibody sequences are numbered according to the Kabat scheme.
Similarly, the terms "CDR", "CDRL1", "CDRL2", "CDRL3", "CDRH1",
"CDRH2", "CDRH3" follow the Kabat numbering system as set forth in
Kabat et al; Sequences of proteins of Immunological Interest NIH,
1987.
[0172] The terms "Variant" refers to at least one, two or three
amino acid changes in the sequence. These amino acid changes may be
deletion, substitution or addition but are preferably substitution.
In one such embodiment the substitutions are conservative
substitutions.
[0173] In an alternative embodiment the variant sequence contains
at least one substitution whilst retaining the canonical of the
antigen binding protein.
[0174] The complementarity determining regions (CDRs) L1, L2, L3,
H1 and H2 tend to structurally exhibit one of a finite number of
main chain conformations. The particular canonical structure class
of a CDR is defined by both the length of the CDR and by the loop
packing, determined by residues located at key positions in both
the CDRs and the framework regions (structurally determining
residues or SDRs). Martin and Thornton (1996; J Mol Biol
263:800-815) have generated an automatic method to define the "key
residue" canonical templates. Cluster analysis is used to define
the canonical classes for sets of CDRs, and canonical templates are
then identified by analysing buried hydrophobics, hydrogen-bonding
residues, and e.g. conserved glycines. The CDRs of antibody
sequences can be assigned to canonical classes by comparing the
sequences to the key residue templates and scoring each template
using identity or similarity matrices.
[0175] The terms "VH" and "VL" are used herein to refer to the
heavy chain variable domain and light chain variable domain
respectively of an antibody.
[0176] As used herein the term "domain" refers to a folded protein
structure which has tertiary structure independent of the rest of
the protein. Generally, domains are responsible for discrete
functional properties of proteins and in many cases may be added,
removed or transferred to other proteins without loss of function
of the remainder of the protein and/or of the domain. An "antibody
single variable domain" is a folded polypeptide domain comprising
sequences characteristic of antibody variable domains. It therefore
includes complete antibody variable domains and modified variable
domains, for example, in which one or more loops have been replaced
by sequences which are not characteristic of antibody variable
domains, or antibody variable domains which have been truncated or
comprise N- or C-terminal extensions, as well as folded fragments
of variable domains which retain at least the binding activity and
specificity of the full-length domain.
[0177] The phrase "immunoglobulin single variable domain" refers to
an antibody variable domain (VH, VHH, VL) that specifically binds
an antigen or epitope independently of a different V region or
domain. An immunoglobulin single variable domain can be present in
a format (e.g., homo- or hetero-multimer) with other, different
variable regions or variable domains where the other regions or
domains are not required for antigen binding by the single
immunoglobulin variable domain (i.e., where the immunoglobulin
single variable domain binds antigen independently of the
additional variable domains). A "domain antibody" or "dAb" is the
same as an "immunoglobulin single variable domain" which is capable
of binding to an antigen as the term is used herein. An
immunoglobulin single variable domain may be a human antibody
variable domain, but also includes single antibody variable domains
from other species such as rodent (for example, as disclosed in WO
00/29004), nurse shark and Camelid VHH dAbs. Camelid VHH are
immunoglobulin single variable domain polypeptides that are derived
from species including camel, llama, alpaca, dromedary, and
guanaco, which produce heavy chain antibodies naturally devoid of
light chains. Such VHH domains may be humanised according to
standard techniques available in the art, and such domains are
still considered to be "domain antibodies" according to the
invention. As used herein "VH includes camelid VHH domains. NARV
are another type of immunoglobulin single variable domain which
were identified in cartilaginous fish including the nurse shark.
These domains are also known as Novel Antigen Receptor variable
region (commonly abbreviated to V(NAR) or NARV). For further
details see Mol. Immunol. 44, 656-665 (2006) and
US20050043519A.
[0178] The term "Epitope-binding domain" refers to a domain that
specifically binds an antigen or epitope independently of a
different V region or domain, this may be a domain antibody (dAb),
for example a human, camelid or shark immunoglobulin single
variable domain or it may be a domain which is a derivative of a
scaffold selected from the group consisting of CTLA-4 (Evibody);
lipocalin; Protein A derived molecules such as Z-domain of Protein
A (Affibody, SpA), A-domain (Avimer/Maxibody); Heat shock proteins
such as GroEl and GroES; 29eroxidise29g (trans-body); ankyrin
repeat protein (DARPin); peptide aptamer; C-type lectin domain
(Tetranectin); human .gamma.-crystallin and human ubiquitin
(affilins); PDZ domains; scorpion toxinkunitz type domains of human
protease inhibitors; and fibronectin (adnectin); which has been
subjected to protein engineering in order to obtain binding to a
ligand other than the natural ligand.
[0179] CTLA-4 (Cytotoxic T Lymphocyte-associated Antigen 4) is a
CD28-family receptor expressed on mainly CD4+ T-cells. Its
extracellular domain has a variable domain-like Ig fold. Loops
corresponding to CDRs of antibodies can be substituted with
heterologous sequence to confer different binding properties.
CTLA-4 molecules engineered to have different binding specificities
are also known as Evibodies. For further details see Journal of
Immunological Methods 248 (1-2), 31-45 (2001)
[0180] Lipocalins are a family of extracellular proteins which
transport small hydrophobic molecules such as steroids, bilins,
retinoids and lipids. They have a rigid .beta.-sheet secondary
structure with a number of loops at the open end of the conical
structure which can be engineered to bind to different target
antigens. Anticalins are between 160-180 amino acids in size, and
are derived from lipocalins. For further details see Biochim
Biophys Acta 1482: 337-350 (2000), U.S. Pat. No. 7,250,297B1 and
US20070224633
[0181] An affibody is a scaffold derived from Protein A of
Staphylococcus aureus which can be engineered to bind to antigen.
The domain consists of a three-helical bundle of approximately 58
amino acids. Libraries have been generated by randomisation of
surface residues. For further details see Protein Eng. Des. Sel.
17, 455-462 (2004) and EP1641818A1
[0182] Avimers are multidomain proteins derived from the A-domain
scaffold family. The native domains of approximately 35 amino acids
adopt a defined disulphide bonded structure. Diversity is generated
by shuffling of the natural variation exhibited by the family of
A-domains. For further details see Nature Biotechnology 23(12),
1556-1561 (2005) and Expert Opinion on Investigational Drugs 16(6),
909-917 (June 2007)
[0183] A Transferrin is a monomeric serum transport glycoprotein.
Transferrins can be engineered to bind different target antigens by
insertion of peptide sequences in a permissive surface loop.
Examples of engineered transferrins scaffolds include the
Trans-body. For further details see J. Biol. Chem 274, 24066-24073
(1999).
[0184] Designed Ankyrin Repeat Proteins (DARPins) are derived from
Ankyrin which is a family of proteins that mediate attachment of
integral membrane proteins to the cytoskeleton. A single ankyrin
repeat is a 33 residue motif consisting of two .alpha.-helices and
a .beta.-turn. They can be engineered to bind different target
antigens by randomising residues in the first .alpha.-helix and a
.beta.-turn of each repeat. Their binding interface can be
increased by increasing the number of modules (a method of affinity
maturation). For further details see J. Mol. Biol. 332, 489-503
(2003), PNAS 100(4), 1700-1705 (2003) and J. Mol. Biol. 369,
1015-1028 (2007) and US20040132028A1.
[0185] Fibronectin is a scaffold which can be engineered to bind to
antigen. Adnectins consists of a backbone of the natural amino acid
sequence of the 10.sup.th domain of the 15 repeating units of human
fibronectin type III (FN3). Three loops at one end of the
.beta.-sandwich can be engineered to enable an Adnectin to
specifically recognize a therapeutic target of interest. For
further details see Protein Eng. Des. Sel. 18, 435-444 (2005),
US20080139791, WO2005056764 and U.S. Pat. No. 6,818,418B1.
[0186] Peptide aptamers are combinatorial recognition molecules
that consist of a constant scaffold protein, typically thioredoxin
(TrxA) which contains a constrained variable peptide loop inserted
at the active site. For further details see Expert Opin. Biol.
Ther. 5, 783-797 (2005).
[0187] Microbodies are derived from naturally occurring
microproteins of 25-50 amino acids in length which contain 3-4
cysteine bridges--examples of microproteins include KalataB1 and
conotoxin and knottins. The microproteins have a loop which can be
engineered to include up to 25 amino acids without affecting the
overall fold of the microprotein. For further details of engineered
knottin domains, see WO2008098796.
[0188] Other epitope binding domains include proteins which have
been used as a scaffold to engineer different target antigen
binding properties include human .gamma.-crystallin and human
ubiquitin (affilins), kunitz type domains of human protease
inhibitors, PDZ-domains of the Ras-binding protein AF-6, scorpion
toxins (charybdotoxin), C-type lectin domain (tetranectins) are
reviewed in Chapter 7--Non-Antibody Scaffolds from Handbook of
Therapeutic Antibodies (2007, edited by Stefan Dubel) and Protein
Science 15:14-27 (2006). Epitope binding domains of the present
invention could be derived from any of these alternative protein
domains.
[0189] As used herein, the term "antigen-binding site" refers to a
site on a protein which is capable of specifically binding to
antigen, this may be a single domain, for example an
epitope-binding domain, or it may be paired VH/VL domains as can be
found on a standard antibody. In some embodiments of the invention
single-chain Fv (ScFv) domains can provide antigen-binding
sites.
[0190] The terms "mAbdAb" and dAbmAb" are used herein to refer to
antigen-binding proteins of the present invention. The two terms
can be used interchangeably, and are intended to have the same
meaning as used herein.
[0191] The term "antigen binding protein" as used herein refers to
antibodies, antibody fragments for example a domain antibody (dAb),
ScFv, Fab, Fab2, and other protein constructs. Antigen binding
molecules may comprise at least one Ig variable domain, for example
antibodies, domain antibodies (dAbs), Fab, Fab', F(ab')2, Fv, ScFv,
diabodies, mAbdAbs, affibodies, heteroconjugate antibodies or
bispecific antibodies. In one embodiment the antigen binding
molecule is an antibody. In another embodiment the antigen binding
molecule is a dAb, i.e. an immunoglobulin single variable domain
such as a VH, VHH or VL that specifically binds an antigen or
epitope independently of a different V region or domain. Antigen
binding molecules may be capable of binding to two targets, i.e.
they may be dual targeting proteins. Antigen binding molecules may
be a combination of antibodies and antigen binding fragments such
as for example, one or more domain antibodies and/or one or more
ScFvs linked to a monoclonal antibody. Antigen binding molecules
may also comprise a non-Ig domain for example a domain which is a
derivative of a scaffold selected from the group consisting of
CTLA-4 (Evibody); lipocalin; Protein A derived molecules such as
Z-domain of Protein A (Affibody, SpA), A-domain (Avimer/Maxibody);
Heat shock proteins such as GroEl and GroES; 32eroxidise32g
(trans-body); ankyrin repeat protein (DARPin); peptide aptamer;
C-type lectin domain (Tetranectin); human .gamma.-crystallin and
human ubiquitin (affilins); PDZ domains; scorpion toxinkunitz type
domains of human protease inhibitors; and fibronectin (adnectin);
which has been subjected to protein engineering in order to obtain
binding to OSM. As used herein "antigen binding protein" will be
capable of antagonising and/or neutralising human OSM. In addition,
an antigen binding protein may inhibit and or block OSM activity by
binding to OSM and preventing a natural ligand from binding and/or
activating the gp130 receptor.
[0192] The term "Effector Function" as used herein is meant to
refer to one or more of Antibody dependant cell mediated cytotoxic
activity (ADCC), Complement-dependant cytotoxic activity (CDC)
mediated responses, Fc-mediated phagocytosis and antibody recycling
via the FcRn receptor. For IgG antibodies, effector functionalities
including ADCC and ADCP are mediated by the interaction of the
heavy chain constant region with a family of Fc.gamma. receptors
present on the surface of immune cells. In humans these include
Fc.gamma.RI (CD64), Fc.gamma.RII (CD32) and Fc.gamma.RIII (CD16).
Interaction between the antigen binding protein bound to antigen
and the formation of the Fc/Fc.gamma. complex induces a range of
effects including cytotoxicity, immune cell activation,
phagocytosis and release of inflammatory cytokines.
[0193] The interaction between the constant region of an antigen
binding protein and various Fc receptors (FcR) is believed to
mediate the effector functions of the antigen binding protein.
Significant biological effects can be a consequence of effector
functionality, in particular, antibody-dependent cellular
cytotoxicity (ADCC), fixation of complement (complement dependent
cytotoxicity or CDC), and half-life/clearance of the antigen
binding protein. Usually, the ability to mediate effector function
requires binding of the antigen binding protein to an antigen and
not all antigen binding proteins will mediate every effector
function.
[0194] Effector function can be measured in a number of ways
including for example via binding of the Fc.gamma.RIII to Natural
Killer cells or via Fc.gamma.RI to monocytes/macrophages to measure
for ADCC effector function. For example an antigen binding protein
of the present invention can be assessed for ADCC effector function
in a Natural Killer cell assay. Examples of such assays can be
found in Shields et al, 2001 The Journal of Biological Chemistry,
Vol. 276, p6591-6604; Chappel et al, 1993 The Journal of Biological
Chemistry, Vol 268, p25124-25131; Lazar et al, 2006 PNAS, 103;
4005-4010.
[0195] Examples of assays to determine CDC function include that
described in 1995 J Imm Meth 184:29-38.
[0196] Some isotypes of human constant regions, in particular IgG4
and IgG2 isotypes, essentially lack the functions of a) activation
of complement by the classical pathway; and b) antibody-dependent
cellular cytotoxicity. Various modifications to the heavy chain
constant region of antigen binding proteins may be carried out
depending on the desired effector property. IgG1 constant regions
containing specific mutations have separately been described to
reduce binding to Fc receptors and therefore reduce ADCC and CDC
(Duncan et al. Nature 1988, 332; 563-564; Lund et al. J. Immunol.
1991, 147; 2657-2662; Chappel et al. PNAS 1991, 88; 9036-9040;
Burton and Woof, Adv. Immunol. 1992, 51; 1-84; Morgan et al.,
Immunology 1995, 86; 319-324; Hezareh et al., J. Virol. 2001, 75
(24); 12161-12168).
[0197] In one embodiment of the present invention there is provided
an antigen binding protein comprising a constant region such that
the antigen binding protein has reduced ADCC and/or complement
activation or effector functionality. In one such embodiment the
heavy chain constant region may comprise a naturally disabled
constant region of IgG2 or IgG4 isotype or a mutated IgG1 constant
region. Examples of suitable modifications are described in
EP0307434. One example comprises the substitutions of alanine
residues at positions 235 and 237 (EU index numbering).
[0198] Human IgG1 constant regions containing specific mutations or
altered glycosylation on residue Asn297 have also been described to
enhance binding to Fc receptors. In some cases these mutations have
also been shown to enhance ADCC and CDC (Lazar et al. PNAS 2006,
103; 4005-4010; Shields et al. J Biol Chem 2001, 276; 6591-6604;
Nechansky et al. Mol Immunol, 2007, 44; 1815-1817).
[0199] In one embodiment of the present invention, such mutations
are in one or more of positions selected from 239, 332 and 330
(IgG1), or the equivalent positions in other IgG isotypes. Examples
of suitable mutations are S239D and 1332E and A330L. In one
embodiment the antigen binding protein of the invention herein
described is mutated at positions 239 and 332, for example S239D
and 1332E or in a further embodiment it is mutated at three or more
positions selected from 239 and 332 and 330, for example S239D and
1332E and A330L. (EU index numbering).
[0200] In an alternative embodiment of the present invention, there
is provided an antigen binding protein comprising a heavy chain
constant region with an altered glycosylation profile such that the
antigen binding protein has enhanced effector function. For
example, wherein the antigen binding protein has enhanced ADCC or
enhanced CDC or wherein it has both enhanced ADCC and CDC effector
function. Examples of suitable methodologies to produce antigen
binding proteins with an altered glycosylation profile are
described in WO2003011878, WO2006014679 and EP1229125, all of which
can be applied to the antigen binding proteins of the present
invention.
[0201] The present invention also provides a method for the
production of an antigen binding protein according to the invention
comprising the steps of:
a) culturing a recombinant host cell comprising an expression
vector comprising the isolated nucleic acid as described herein,
wherein the FUT8 gene encoding alpha-1,6-fucosyltransferase has
been inactivated in the recombinant host cell; and b) recovering
the antigen binding protein.
[0202] Such methods for the production of antigen binding proteins
can be performed, for example, using the POTELLIGENT.TM. technology
system available from BioWa, Inc. (Princeton, N.J.) in which
CHOK1SV cells lacking a functional copy of the FUT8 gene produce
monoclonal antibodies having enhanced antibody dependent cell
mediated cytotoxicity (ADCC) activity that is increased relative to
an identical monoclonal antibody produced in a cell with a
functional FUT8 gene. Aspects of the POTELLIGENT.TM. technology
system are described in U.S. Pat. No. 7,214,775, U.S. Pat. No.
6,946,292, WO0061739 and WO0231240 all of which are incorporated
herein by reference. Those of ordinary skill in the art will also
recognize other appropriate systems.
[0203] In one embodiment of the present invention there is provided
an antigen binding protein comprising a chimaeric heavy chain
constant region for example an antigen binding protein comprising a
chimaeric heavy chain constant region with at least one CH2 domain
from IgG3 such that the antigen binding protein has enhanced
effector function, for example wherein it has enhanced ADCC or
enhanced CDC, or enhanced ADCC and CDC functions. In one such
embodiment, the antigen binding protein may comprise one CH2 domain
from IgG3 or both CH2 domains may be from IgG3.
[0204] Also provided is a method of producing an antigen binding
protein according to the invention comprising the steps of:
a) culturing a recombinant host cell comprising an expression
vector comprising an isolated nucleic acid as described herein
wherein the expression vector comprises a nucleic acid sequence
encoding an Fc domain having both IgG1 and IgG3 Fc domain amino
acid residues; and b) recovering the antigen binding protein.
[0205] Such methods for the production of antigen binding proteins
can be performed, for example, using the COMPLEGENT.TM. technology
system available from BioWa, Inc. (Princeton, N.J.) and Kyowa Hakko
Kogyo (now, Kyowa Hakko Kirin Co., Ltd.) Co., Ltd. In which a
recombinant host cell comprising an expression vector in which a
nucleic acid sequence encoding a chimeric Fc domain having both
IgG1 and IgG3 Fc domain amino acid residues is expressed to produce
an antigen binding protein having enhanced complement dependent
cytotoxicity (CDC) activity that is increased relative to an
otherwise identical antigen binding protein lacking such a chimeric
Fc domain. Aspects of the COMPLEGENT.TM. technology system are
described in WO2007011041 and US20070148165 each of which are
incorporated herein by reference. In an alternative embodiment CDC
activity may be increased by introducing sequence specific
mutations into the Fc region of an IgG chain. Those of ordinary
skill in the art will also recognize other appropriate systems.
[0206] It will be apparent to those skilled in the art that such
modifications may not only be used alone but may be used in
combination with each other in order to further enhance effector
function. In one such embodiment of the present invention there is
provided an antigen binding protein comprising a heavy chain
constant region which comprises a mutated and chimaeric heavy chain
constant region for example wherein an antigen binding protein
comprising at least one CH2 domain from IgG3 and one CH2 domain
from IgG1, wherein the IgG1 CH2 domain has one or more mutations at
positions selected from 239 and 332 and 330 (for example the
mutations may be selected from S239D and 1332E and A330L) such that
the antigen binding protein has enhanced effector function, for
example wherein it has one or more of the following functions,
enhanced ADCC or enhanced CDC, for example wherein it has enhanced
ADCC and enhanced CDC. In one embodiment the IgG1 CH2 domain has
the mutations S239D and 1332E.
[0207] In an alternative embodiment of the present invention there
is provided an antigen binding protein comprising a chimaeric heavy
chain constant region and which has an altered glycosylation
profile. In one such embodiment the heavy chain constant region
comprises at least one CH2 domain from IgG3 and one CH2 domain from
IgG1 and has an altered glycosylation profile such that the ratio
of fucose to mannose is 0.8:3 or less, for example wherein the
antigen binding protein is defucosylated so that said antigen
binding protein has an enhanced effector function in comparison
with an equivalent antigen binding protein with an immunoglobulin
heavy chain constant region lacking said mutations and altered
glycosylation profile, for example wherein it has one or more of
the following functions, enhanced ADCC or enhanced CDC, for example
wherein it has enhanced ADCC and enhanced CDC In an alternative
embodiment the antigen binding protein has at least one IgG3 CH2
domain and at least one heavy chain constant domain from IgG1
wherein both IgG CH2 domains are mutated in accordance with the
limitations described herein.
[0208] In one aspect of the invention there is provided a method of
producing an antigen binding protein according to the invention
described herein comprising the steps of:
a) culturing a recombinant host cell containing an expression
vector containing an isolated nucleic acid as described herein,
said expression vector further comprising a Fc nucleic acid
sequence encoding a chimeric Fc domain having both IgG1 and IgG3 Fc
domain amino acid residues, and wherein the FUT8 gene encoding
alpha-1,6-fucosyltransferase has been inactivated in the
recombinant host cell; and b) recovering the antigen binding
protein.
[0209] Such methods for the production of antigen binding proteins
can be performed, for example, using the ACCRETAMAB.TM. technology
system available from BioWa, Inc. (Princeton, N.J.) which combines
the POTELLIGENT.TM. and COMPLEGENT.TM. technology systems to
produce an antigen binding protein having both ADCC and CDC
enhanced activity that is increased relative to an otherwise
identical monoclonal antibody lacking a chimeric Fc domain and
which has fucose on the oligosaccharide
[0210] In yet another embodiment of the present invention there is
provided an antigen binding protein comprising a mutated and
chimeric heavy chain constant region wherein said antigen binding
protein has an altered glycosylation profile such that the antigen
binding protein has enhanced effector function, for example wherein
it has one or more of the following functions, enhanced ADCC or
enhanced CDC. In one embodiment the mutations are selected from
positions 239 and 332 and 330, for example the mutations are
selected from S239D and I332E and A330L. In a further embodiment
the heavy chain constant region comprises at least one CH2 domain
from IgG3 and one Ch2 domain from IgG1. In one embodiment the heavy
chain constant region has an altered glycosylation profile such
that the ratio of fucose to mannose is 0.8:3 or less for example
the antigen binding protein is defucosylated, so that said antigen
binding protein has an enhanced effector function in comparison
with an equivalent non-chimaeric antigen binding protein or with an
immunoglobulin heavy chain constant region lacking said mutations
and altered glycosylation profile.
Immunoconjugates
[0211] Also provided is an immunoconjugate (interchangeably
referred to as "antibody-drug conjugates," or "ADCs") comprising an
antigen binding protein according to the invention as herein
described including, but not limited to, an antibody conjugated to
one or more cytotoxic agents, such as a chemotherapeutic agent, a
drug, a growth inhibitory agent, a toxin (e.g., a protein toxin, an
enzymatically active toxin of bacterial, fungal, plant, or animal
origin, or fragments thereof), or a radioactive isotope (i.e., a
radioconjugate).
[0212] Immunoconjugates have been used for the local delivery of
cytotoxic agents, i.e., drugs that kill or inhibit the growth or
proliferation of cells, in the treatment of cancer (Lambert, J.
(2005) Curr. Opinion in Pharmacology 5:543-549; Wu et al. (2005)
Nature Biotechnology 23(9):1137-1146; Payne, G. (2003) i 3:207-212;
Syrigos and Epenetos (1999) Anticancer Research 19:605-614;
Niculescu-Duvaz and Springer (1997) Adv. Drug Deliv. Rev.
26:151-172; U.S. Pat. No. 4,975,278). Immunoconjugates allow for
the targeted delivery of a drug moiety to a tumor, and
intracellular accumulation therein, where systemic administration
of unconjugated drugs may result in unacceptable levels of toxicity
to normal cells as well as the tumor cells sought to be eliminated
(Baldwin et al., Lancet (Mar. 15, 1986) pp. 603-05; Thorpe (1985)
"Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A
Review," in Monoclonal Antibodies '84: Biological And Clinical
Applications (A. Pinchera et al., eds) pp. 475-506. Both polyclonal
antibodies and monoclonal antibodies have been reported as useful
in these strategies (Rowland et al., (1986) Cancer Immunol.
Immunother. 21:183-87). Drugs used in these methods include
daunomycin, doxorubicin, methotrexate, and vindesine (Rowland et
al., (1986) supra). Toxins used in antibody-toxin conjugates
include bacterial toxins such as diphtheria toxin, plant toxins
such as ricin, small molecule toxins such as geldanamycin (Mandler
et al (2000) J. Nat. Cancer Inst. 92(19):1573-1581; Mandler et al
(2000) Bioorganic & Med. Chem. Letters 10:1025-1028; Mandler et
al (2002) Bioconjugate Chem. 13:786-791), maytansinoids (EP
1391213; Liu et al., (1996) Proc. Natl. Acad. Sci. USA
93:8618-8623), and calicheamicin (Lode et al (1998) Cancer Res.
58:2928; Hinman et al (1993) Cancer Res. 53:3336-3342).
[0213] In one embodiment, the present invention includes
immunoconjugates having the following general structure:
ABP-((Linker).sub.n-Ctx).sub.m
Wherein ABP is an antigen binding protein Linker is either absent
or any a cleavable or non-cleavable linker described herein Ctx is
any cytotoxic agent described herein n is 0, 1, 2, or 3 and m is 1,
2, 3, 4, 5, 6, 7, 8, 9 or 10.
[0214] Examples of antibodies linked by an MC linker with
auristatins such as MMAE and MMAF are depicted in the following
structures:
##STR00001##
[0215] In certain embodiments, an immunoconjugate comprises an
antigen binding protein, including but not limited to, an antibody
and a chemotherapeutic agent or other toxin. Chemotherapeutic
agents useful in the generation of immunoconjugates are described
herein. Enzymatically active toxins and fragments thereof that can
be used include diphtheria A chain, nonbinding active fragments of
diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa),
ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin,
Aleurites fordii proteins, dianthin proteins, Phytolaca americana
proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor,
curcin, crotin, sapaonaria officinalis inhibitor, gelonin,
mitogellin, restrictocin, phenomycin, enomycin, and the
tricothecenes. See, e.g., WO 93/21232 published Oct. 28, 1993. A
variety of radionuclides are available for the production of
radioconjugated antibodies. Examples include .sup.211At,
.sup.212Bi, .sup.131I, .sup.131In, .sup.90Y, and .sup.186Re.
[0216] Antigen binding proteins of the present invention may also
be conjugated to one or more toxins, including, but not limited to,
a calicheamicin, maytansinoids, dolastatins, aurostatins, a
trichothecene, and CC1065, and the derivatives of these toxins that
have toxin activity. Suitable cytotoxic agents include, but are not
limited to, an auristatin including
dovaline-valine-dolaisoleunine-dolaproine-phenylalanine (MMAF) and
monomethyl auristatin E (MMAE) as well as ester forms of MMAE, a
DNA minor groove binding agent, a DNA minor groove alkylating
agent, an enediyne, a lexitropsin, a duocarmycin, a taxane,
including paclitaxel and docetaxel, a puromycin, a dolastatin, a
maytansinoid, and a vinca alkaloid. Specific cytotoxic agents
include topotecan, morpholino-doxorubicin, rhizoxin,
cyanomorpholino-doxorubicin, dolastatin-10, echinomycin,
combretatstatin, chalicheamicin, maytansine, DM-1, DM-4, netropsin.
Other suitable cytotoxic agents include anti-tubulin agents, such
as an auristatin, a vinca alkaloid, a podophyllotoxin, a taxane, a
baccatin derivative, a cryptophysin, a maytansinoid, a
combretastatin, or a dolastatin. Antitubulin agent include
dimethylvaline-valine-dolaisoleuine-dolaproine-phenylalanine-p-ph-
enylened-iamine (AFP), MMAF, MMAE, auristatin E, vincristine,
vinblastine, vindesine, vinorelbine, VP-16, camptothecin,
paclitaxel, docetaxel, epothilone A, epothilone B, nocodazole,
colchicines, colcimid, estramustine, cemadotin, discodermolide,
maytansine, DM-1, DM-4 or eleutherobin.
[0217] Antibody drug conjugates were produced by conjugating the
small molecule anti-tubulin agent monomethylauristatin E (MMAE) or
monomethylauristatin F (MMAF) to the antibodies. In the case of
MMAE the linker consists of a thiol-reactive maleimide, a caproyl
spacer, the dipeptide valine-citrulline, and
p-aminobenzyloxycarbonyl, a self-immolative fragmenting group. In
the case of MMAF a protease-resistant maleimidocaproyl linker is
used. The conjugation process leads to heterogeneity in
drug-antibody attachment, varying in both the number of drugs bound
to each antibody molecule (mole ratio [MR]), and the site of
attachment. The most prevalent species is the material with an
MR=4; less prevalent are materials with MR of 0, 2, 6, and 8. The
overall average drug-to-antibody MR is approximately 4.
Production of Immunoconjugates
[0218] The points of attachment are cysteines produced by mild
reduction of the interchain disulfides of the antibody which is
carried out whilst antibodies are immobilised on Protein G affinity
resin (thus enabling the use of large reagent excesses without
intermediate purifications). While immobilized, a large excess of
TCEP will fully reduce the interchain disulfides but has no impact
upon the binding of the antibody to the resin. [0219] The number of
thiols per antibody generated by this procedure depends upon the
source and isotype of the antibodies. For example, human (and
mouse-human chimeric) IgG1s have 4 reducible disulfides, and thus
generate 8 thiols upon full reduction, whereas murine IgG1s have 5
reducible disulfides and produce 10 thiols. If ADCs with the
maximal drug loading (e.g., 10 drugs per antibody for the murine
IgG1s) are desired, then the maleimido-drug-linker can simply be
added to the immobilized antibodies in sufficient excess to ensure
complete conjugation. However, ADCs with fewer drugs per antibody
can also be prepared from fully reduced antibodies by including a
biologically inert capping agent such as N-ethyl maleimide (NEM)
which occupies some of the available thiols on the antibody. When
the maleimido-drug-linker and the capping agent are added
simultaneously to the fully reduced antibody and in large excess
(at least 3-fold), the two maleimide electrophiles compete for the
limiting number of available thiols. In this fashion, the drug
loading is determined by the relative thiol reaction rates of the
drug-linker and capping agent, and thus can be considered to be
under kinetic control. The relative reaction rates of
maleimido-drug-linkers do vary significantly, and thus the molar
ratio of drug-linker to NEM present in a reaction mix must be
determined empirically to arrive at a panel of ADCs with a desired
level of drug loading. The mole fraction of the drug linkers
SGD-1006 (vcMMAE) and SGD-1269 (mcMMAF) in NEM mixtures which yield
ADCs with approximately 4 drugs per antibody are summarized in
Table 2 for common human and murine IgG isotypes.
Auristatins and Dolastatins
[0220] In some embodiments, the immunoconjugate comprises an
antigen binding protein or antibody conjugated to dolastatins or
dolostatin peptidic analogs and derivatives, the auristatins (U.S.
Pat. Nos. 5,635,483; 5,780,588). Dolastatins and auristatins have
been shown to interfere with microtubule dynamics, GTP hydrolysis,
and nuclear and cellular division (Woyke et al. (2001) Antimicrob.
Agents and Chemother. 45(12):3580-3584) and have anticancer (U.S.
Pat. No. 5,663,149) and antifungal activity (Pettit et al. (1998)
Antimicrob. Agents Chemother. 42:2961-2965). The dolastatin or
auristatin (which are pentapeptide derivatives of dolastatins) drug
moiety may be attached to the antibody through the N (amino)
terminus or the C (carboxyl) terminus of the peptidic drug moiety
(WO 02/088172).
[0221] Exemplary auristatin embodiments include the N-terminus
linked monomethylauristatin drug moieties DE and DF, disclosed in
"Monomethylvaline Compounds Capable of Conjugation to Ligands,"
U.S. Pat. No. 7,498,298, the disclosure of which is expressly
incorporated by reference in its entirety. As used herein, the
abbreviation "MMAE" refers to monomethyl auristatin E. As used
herein the abbreviation "MMAF" refers to
dovaline-valine-dolaisoleuine-dolaproine-phenylalanine.
[0222] Typically, peptide-based drug moieties can be prepared by
forming a peptide bond between two or more amino acids and/or
peptide fragments. Such peptide bonds can be prepared, for example,
according to the liquid phase synthesis method (see E. Schroder and
K. Lubke, "The Peptides," volume 1, pp 76-136, 1965, Academic
Press) that is well known in the field of peptide chemistry. The
auristatin/dolastatin drug moieties may be prepared according to
the methods of: U.S. Pat. No. 5,635,483; U.S. Pat. No. 5,780,588;
Pettit et al. (1989) J. Am. Chem. Soc. 111:5463-5465; Pettit et al.
(1998) Anti-Cancer Drug Design 13:243-277; Pettit, G. R., et al.
Synthesis, 1996, 719-725; and Pettit et al. (1996) J. Chem. Soc.
Perkin Trans. 15:859-863. See also Doronina (2003) Nat Biotechnol
21(7):778-784; "Monomethylvaline Compounds Capable of Conjugation
to Ligands," U.S. Pat. No. 7,498,298, filed Nov. 5, 2004, hereby
incorporated by reference in its entirety (disclosing, e.g.,
linkers and methods of preparing monomethylvaline compounds such as
MMAE and MMAF conjugated to linkers). Biologically active organic
compounds which act as cytotoxic agents, specifically
pentapeptides, are disclosed in U.S. Pat. Nos. 6,884,869;
7,498,298; 7,098,308; 7,256,257; and 7,423,116. Monoclonal
antibodies linked with MMAE and MMAF as well as various derivatives
of auristatins and methods of making them are described in U.S.
Pat. No. 7,964,566.
[0223] Examples of auristatins include MMAE and MMAF the structures
of which are shown below:
##STR00002##
Maytansine and Maytansinoids
[0224] Maytansinoids are mitototic inhibitors which act by
inhibiting tubulin polymerization. Maytansine was first isolated
from the east African shrub Maytenus serrata (U.S. Pat. No.
3,896,111). Subsequently, it was discovered that certain microbes
also produce maytansinoids, such as maytansinol and C-3 maytansinol
esters (U.S. Pat. No. 4,151,042). Highly cytotoxic maytansinoid
drugs can be prepared from ansamitocin precursors produced by
fermentation of microorganisms such as Actinosynnema. Methods for
isolating ansamitocins are described in U.S. Pat. No. 6,573,074.
Synthetic maytansinol and derivatives and analogues thereof are
disclosed, for example, in U.S. Pat. Nos. 4,137,230; 4,248,870;
4,256,746; 4,260,608; 4,265,814; 4,294,757; 4,307,016; 4,308,268;
4,308,269; 4,309,428; 4,313,946; 4,315,929; 4,317,821; 4,322,348;
4,331,598; 4,361,650; 4,364,866; 4,424,219; 4,450,254; 4,362,663;
and 4,371,533.
[0225] Antibody-maytansinoid conjugates are prepared by chemically
linking an antibody to a maytansinoid molecule without
significantly diminishing the biological activity of either the
antibody or the maytansinoid molecule. See, e.g., U.S. Pat. No.
5,208,020. An average of 3-4 maytansinoid molecules conjugated per
antibody molecule has shown efficacy in enhancing cytotoxicity of
target cells without negatively affecting the function or
solubility of the antibody, although even one molecule of
toxin/antibody would be expected to enhance cytotoxicity over the
use of naked antibody. Maytansinoids are well known in the art and
can be synthesized by known techniques or isolated from natural
sources. Suitable maytansinoids are disclosed, for example, in U.S.
Pat. No. 5,208,020 and in the other patents and nonpatent
publications referred to hereinabove. Maytansinoids are maytansinol
and maytansinol analogues modified in the aromatic ring or at other
positions of the maytansinol molecule, such as various maytansinol
esters. Methods for preparing matansinoids for linkage with
antibodies are disclosed in U.S. Pat. Nos. 6,570,024 and
6,884,874.
Calicheamicin
[0226] The calicheamicin family of antibiotics is capable of
producing double-stranded DNA breaks at sub-picomolar
concentrations. For the preparation of conjugates of the
calicheamicin family, see U.S. Pat. Nos. 5,712,374, 5,714,586,
5,739,116, 5,767,285, 5,770,701, 5,770,710, 5,773,001, 5,877,296
(all to American Cyanamid Company). Structural analogues of
calicheamicin which may be used include, but are not limited to,
.gamma.1I, .alpha.2I, .alpha.3I, N-acetyl-.gamma.1I, PSAG and
.theta.I1 (Hinman et al., Cancer Research 53:3336-3342 (1993), Lode
et al., Cancer Research 58:2925-2928 (1998) and the aforementioned
U.S. patents to American Cyanamid). Another anti-tumor drug that
the antibody can be conjugated is QFA which is an antifolate. Both
calicheamicin and QFA have intracellular sites of action and do not
readily cross the plasma membrane. Therefore, cellular uptake of
these agents through antibody mediated internalization greatly
enhances their cytotoxic effects.
Other Cytotoxic Agents
[0227] Other antitumor agents that can be conjugated to the
antibodies include BCNU, streptozoicin, vincristine and
5-fluorouracil, the family of agents known collectively LL-E33288
complex described in U.S. Pat. Nos. 5,053,394, 5,770,710, as well
as esperamicins (U.S. Pat. No. 5,877,296).
[0228] Enzymatically active toxins and fragments thereof which can
be used include diphtheria A chain, nonbinding active fragments of
diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa),
ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin,
Aleurites fordii proteins, dianthin proteins, Phytolaca americana
proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor,
curcin, crotin, sapaonaria officinalis inhibitor, gelonin,
mitogellin, restrictocin, phenomycin, enomycin and the
tricothecenes. See, for example, WO 93/21232 published Oct. 28,
1993.
[0229] The present invention further contemplates an
immunoconjugate formed between an antibody and a compound with
nucleolytic activity (e.g., a ribonuclease or a DNA endonuclease
such as a deoxyribonuclease; DNase).
[0230] For selective destruction of the tumor, the antibody may
comprise a highly radioactive atom. A variety of radioactive
isotopes are available for the production of radioconjugated
antibodies. Examples include At211, I131, I125, Y90, Re186, Re188,
Sm153, Bi212, P32, Pb212 and radioactive isotopes of Lu. When the
conjugate is used for detection, it may comprise a radioactive atom
for scintigraphic studies, for example tc99m or I123, or a spin
label for nuclear magnetic resonance (NMR) imaging (also known as
magnetic resonance imaging, mri), such as iodine-123 again,
iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15,
oxygen-17, gadolinium, manganese or iron.
[0231] The radio- or other labels may be incorporated in the
conjugate in known ways. For example, the peptide may be
biosynthesized or may be synthesized by chemical amino acid
synthesis using suitable amino acid precursors involving, for
example, fluorine-19 in place of hydrogen. Labels such as tc99m or
I123, Re186, Re188 and In111 can be attached via a cysteine residue
in the peptide. Yttrium-90 can be attached via a lysine residue.
The IODOGEN method (Fraker et al. (1978) Biochem. Biophys. Res.
Commun. 80: 49-57) can be used to incorporate iodine-123.
"Monoclonal Antibodies in Immunoscintigraphy" (Chatal, CRC Press
1989) describes other methods in detail.
Preparation of ADCs
[0232] In antibody drug conjugates, the antibody can be conjugated
directly to the cytotoxic agent or via a linker. Suitable linkers
include, for example, cleavable and non-cleavable linkers. A
cleavable linker is typically susceptible to cleavage under
intracellular conditions. Suitable cleavable linkers include, for
example, a peptide linker cleavable by an intracellular protease,
such as lysosomal protease or an endosomal protease. In exemplary
embodiments, the linker can be a dipeptide linker, such as a
valine-citrulline (val-cit) or a phenylalanine-lysine (phe-lys)
linker. Other suitable linkers include linkers hydrolyzable at a pH
of less than 5.5, such as a hydrazone linker. Additional suitable
cleavable linkers include disulfide linkers.
[0233] Bristol-Myers Squibb has described particular lysosomal
enzyme-cleavable antitumor drug conjugates. See, for example, U.S.
Pat. No. 6,214,345. Seattle Genetics has published applications
U.S. Pat. Appl. 2003/0096743 and U.S. Pat. Appl. 2003/0130189,
which describe p-aminobenzylethers in drug delivery agents. The
linkers described in these applications are limited to aminobenzyl
ether compositions.
[0234] Conjugates of the antigen binding protein and cytotoxic
agent may be made using a variety of bifunctional protein coupling
agents such as N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP),
succinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate
(SMCC), iminothiolane (IT), bifunctional derivatives of imidoesters
(such as dimethyl adipimidate HCl), active esters (such as
disuccinimidyl suberate), aldehydes (such as glutaraldehyde),
bis-azido compounds (such as bis(p-azidobenzoyl) hexanediamine),
bis-diazonium derivatives (such as
bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as
toluene 2,6-diisocyanate), and bis-active fluorine compounds (such
as 1,5-difluoro-2,4-dinitrobenzene).
[0235] Additionally the linker may be composed of one or more
linker components. Exemplary linker components include
6-maleimidocaproyl ("MC"), maleimidopropanoyl ("MP"),
valine-citrulline ("val-cit"), alanine-phenylalanine ("ala-phe"),
p-aminobenzyloxycarbonyl ("PAB"), N-Succinimidyl
4-(2-pyridylthio)pentanoate ("SPP"), N-Succinimidyl
4-(N-maleimidomethyl)cyclohexane-1 carboxylate ("SMCC"), and
N-Succinimidyl (4-iodo-acetyl)aminobenzoate ("SIAB"). Additional
linker components are known in the art and some are described
herein. See also "Monomethylvaline Compounds Capable of Conjugation
to Ligands," U.S. Pat. No. 7,498,298, filed Nov. 5, 2004, the
contents of which are hereby incorporated by reference in its
entirety.
[0236] Linkers may also comprises amino acids and/or amino acid
analogs. Amino acid linker components include a dipeptide, a
tripeptide, a tetrapeptide or a pentapeptide. Exemplary dipeptides
include: valine-citrulline (vc or val-cit), alanine-phenylalanine
(af or ala-phe). Exemplary tripeptides include:
glycine-valine-citrulline (gly-val-cit) and glycine-glycine-glycine
(gly-gly-gly). Amino acid residues which comprise an amino acid
linker component include those occurring naturally, as well as
minor amino acids and non-naturally occurring amino acid analogs,
such as citrulline. Amino acid linker components can be designed
and optimized in their selectivity for enzymatic cleavage by a
particular enzyme, for example, a tumor-associated protease,
cathepsin B, C and D, or a plasmin protease.
[0237] Antigen binding proteins and antibodies may be made reactive
for conjugation with linker reagents. Nucleophilic groups on
antibodies include, but are not limited to: (i) N-terminal amine
groups, (ii) side chain amine groups, e.g., lysine, (iii) side
chain thiol groups, e.g. cysteine, and (iv) sugar hydroxyl or amino
groups where the antibody is glycosylated. Amine, thiol, and
hydroxyl groups are nucleophilic and capable of reacting to form
covalent bonds with electrophilic groups on linker moieties and
linker reagents including: (i) active esters such as NHS esters,
HOBt esters, haloformates, and acid halides; (ii) alkyl and benzyl
halides such as haloacetamides; (iii) aldehydes, ketones, carboxyl,
and maleimide groups. Certain antibodies have reducible interchain
disulfides, i.e. cysteine bridges. Antibodies may be made reactive
for conjugation with linker reagents by treatment with a reducing
agent such as DTT (dithiothreitol). Each cysteine bridge will thus
form, theoretically, two reactive thiol nucleophiles. Additional
nucleophilic groups can be introduced into antibodies through the
reaction of lysines with 2-iminothiolane (Traut's reagent)
resulting in conversion of an amine into a thiol. Reactive thiol
groups may be introduced into the antibody (or fragment thereof) by
introducing one, two, three, four, or more cysteine residues (e.g.,
preparing mutant antibodies comprising one or more non-native
cysteine amino acid residues).
[0238] Antigen binding proteins and antibodies may also be modified
to introduce electrophilic moieties, which can react with
nucleophilic substituents on the linker reagent or drug. The sugars
of glycosylated antibodies may be oxidized, e.g. with periodate
oxidizing reagents, to form aldehyde or ketone groups which may
react with the amine group of linker reagents or drug moieties. The
resulting imine Schiff base groups may form a stable linkage, or
may be reduced, e.g., by borohydride reagents to form stable amine
linkages. In one embodiment, reaction of the carbohydrate portion
of a glycosylated antibody with either glactose oxidase or sodium
meta-periodate may yield carbonyl (aldehyde and ketone) groups in
the protein that can react with appropriate groups on the drug
(Hermanson, Bioconjugate Techniques). In another embodiment,
proteins containing N-terminal serine or threonine residues can
react with sodium meta-periodate, resulting in production of an
aldehyde in place of the first amino acid (Geoghegan & Stroh,
(1992) Bioconjugate Chem. 3:138-146; U.S. Pat. No. 5,362,852). Such
aldehydes can be reacted with a drug moiety or linker
nucleophile.
[0239] Nucleophilic groups on a drug moiety include, but are not
limited to: amine, thiol, hydroxyl, hydrazide, oxime, hydrazine,
thiosemicarbazone, hydrazine carboxylate, and arylhydrazide groups
capable of reacting to form covalent bonds with electrophilic
groups on linker moieties and linker reagents including: (i) active
esters such as NHS esters, HOBt esters, haloformates, and acid
halides; (ii) alkyl and benzyl halides such as haloacetamides;
(iii) aldehydes, ketones, carboxyl, and maleimide groups.
[0240] In some embodiments, the linker is cleavable by a cleaving
agent that is present in the intracellular environment (e.g.,
within a lysosome or endosome or caveolea). The linker can be,
e.g., a peptidyl linker that is cleaved by an intracellular
peptidase or protease enzyme, including, but not limited to, a
lysosomal or endosomal protease. Typically, the peptidyl linker is
at least two amino acids long or at least three amino acids long.
Cleaving agents can include cathepsins B and D and plasmin, all of
which are known to hydrolyze dipeptide drug derivatives resulting
in the release of active drug inside target cells (see, e.g.,
Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123).
Peptidyl linkers may be cleavable by enzymes that are present
cells. For example, a peptidyl linker that is cleavable by the
thiol-dependent protease cathepsin-B, which is highly expressed in
cancerous tissue, can be used (e.g., a Phe-Leu or a Gly-Phe-Leu-Gly
(SEQ ID NO:50) linker). Other such linkers are described, e.g., in
U.S. Pat. No. 6,214,345. In specific embodiments, the peptidyl
linker cleavable by an intracellular protease is a Val-Cit linker
or a Phe-Lys linker (see, e.g., U.S. Pat. No. 6,214,345, which
describes the synthesis of doxorubicin with the val-cit linker).
One advantage of using intracellular proteolytic release of the
therapeutic agent is that the agent is typically attenuated when
conjugated and the serum stabilities of the conjugates are
typically high.
[0241] In other embodiments, the cleavable linker is pH-sensitive,
i.e., sensitive to hydrolysis at certain pH values. Typically, the
pH-sensitive linker hydrolyzable under acidic conditions. For
example, an acid-labile linker that is hydrolyzable in the lysosome
(e.g., a hydrazone, semicarbazone, thiosemicarbazone, cis-aconitic
amide, orthoester, acetal, ketal, or the like) can be used. (See,
e.g., U.S. Pat. Nos. 5,122,368; 5,824,805; 5,622,929; Dubowchik and
Walker, 1999, Pharm. Therapeutics 83:67-123; Neville et al., 1989,
Biol. Chem. 264:14653-14661.) Such linkers are relatively stable
under neutral pH conditions, such as those in the blood, but are
unstable at below pH 5.5 or 5.0, the approximate pH of the
lysosome. In certain embodiments, the hydrolyzable linker is a
thioether linker (such as, e.g., a thioether attached to the
therapeutic agent via an acylhydrazone bond (see, e.g., U.S. Pat.
No. 5,622,929)).
[0242] In yet other embodiments, the linker is cleavable under
reducing conditions (e.g., a disulfide linker). A variety of
disulfide linkers are known in the art, including, for example,
those that can be formed using SATA
(N-succinimidyl-5-acetylthioacetate), SPDP
(N-succinimidyl-3-(2-pyridyldithio)propionate), SPDB
(N-succinimidyl-3-(2-pyridyldithio)butyrate) and SMPT
(N-succinimidyl-oxycarbonyl-alpha-methyl-alpha-(2-pyridyl-dithio)toluene)-
-, SPDB and SMPT (See, e.g., Thorpe et al., 1987, Cancer Res.
47:5924-5931; Wawrzynczak et al., In Immunoconjugates: Antibody
Conjugates in Radioimagery and Therapy of Cancer (C. W. Vogel ed.,
Oxford U. Press, 1987. See also U.S. Pat. No. 4,880,935.)
[0243] In yet other specific embodiments, the linker is a malonate
linker (Johnson et al., 1995, Anticancer Res. 15:1387-93), a
maleimidobenzoyl linker (Lau et al., 1995, Bioorg-Med-Chem.
3(10):1299-1304), or a 3'-N-amide analog (Lau et al., 1995,
Bioorg-Med-Chem. 3(10):1305-12).
[0244] Typically, the linker is not substantially sensitive to the
extracellular environment. As used herein, "not substantially
sensitive to the extracellular environment," in the context of a
linker, means that no more than about 20%, typically no more than
about 15%, more typically no more than about 10%, and even more
typically no more than about 5%, no more than about 3%, or no more
than about 1% of the linkers, in a sample of ADC or ADC derivative,
are cleaved when the ADC or ADC derivative present in an
extracellular environment (e.g., in plasma). Whether a linker is
not substantially sensitive to the extracellular environment can be
determined, for example, by incubating independently with plasma
both (a) the ADC or ADC derivative (the "ADC sample") and (b) an
equal molar amount of unconjugated antibody or therapeutic agent
(the "control sample") for a predetermined time period (e.g., 2, 4,
8, 16, or 24 hours) and then comparing the amount of unconjugated
antibody or therapeutic agent present in the ADC sample with that
present in control sample, as measured, for example, by high
performance liquid chromatography.
[0245] In other, non-mutually exclusive embodiments, the linker
promotes cellular internalization. In certain embodiments, the
linker promotes cellular internalization when conjugated to the
therapeutic agent (i.e., in the milieu of the linker-therapeutic
agent moiety of the ADC or ADC derivate as described herein). In
yet other embodiments, the linker promotes cellular internalization
when conjugated to both the therapeutic agent and the antigen
binding protein or antibody or derivative thereof (i.e., in the
milieu of the ADC or ADC derivative as described herein).
[0246] A variety of linkers that can be used with the present
compositions and methods are described in WO 2004010957 entitled
"Drug Conjugates and Their Use for Treating Cancer, An Autoimmune
Disease or an Infectious Disease" filed Jul. 31, 2003, and U.S.
Provisional Application No. 60/400,403, entitled "Drug Conjugates
and their use for treating cancer, an autoimmune disease or an
infectious disease", filed Jul. 31, 2002 (the disclosure of which
is incorporated by reference herein).
[0247] Alternatively, a fusion protein comprising the antigen
binding protein and cytotoxic agent may be made, e.g., by
recombinant techniques or peptide synthesis. The length of DNA may
comprise respective regions encoding the two portions of the
conjugate either adjacent one another or separated by a region
encoding a linker peptide which does not destroy the desired
properties of the conjugate.
[0248] In yet another embodiment, the antibody may be conjugated to
a "receptor" (such as streptavidin) for utilization in tumor
pre-targeting wherein the antibody-receptor conjugate is
administered to the patient, followed by removal of unbound
conjugate from the circulation using a clearing agent and then
administration of a "ligand" (e.g., avidin) which is conjugated to
a cytotoxic agent (e.g., a radionucleotide).
[0249] The term "Non Human antibody or antibody fragment thereof"
as used herein is meant to refer to antibodies or fragments thereof
which originate from any species other than human wherein human
includes chimeric antibodies.
[0250] The term "donor antibody" refers to an antibody (monoclonal,
and/or recombinant) which contributes the amino acid sequences of
its variable domains, CDRs, or other functional fragments or
analogs thereof to a first immunoglobulin partner, so as to provide
the altered immunoglobulin coding region and resulting expressed
altered antibody with the antigenic specificity and neutralizing
activity characteristic of the donor antibody.
[0251] The term "acceptor antibody" refers to an antibody
(monoclonal and/or recombinant) heterologous to the donor antibody,
which contributes all (or any portion, but preferably all) of the
amino acid sequences encoding its heavy and/or light chain
framework regions and/or its heavy and/or light chain constant
regions to the first immunoglobulin partner. The human antibody is
the acceptor antibody. The term "Human acceptor sequence" as used
herein is meant to refer to a framework of an antibody or antibody
fragment thereof comprising the amino acid sequence of a VH or VL
framework derived from a human antibody or antibody fragment
thereof or a human consensus sequence framework into which CDR's
from a non-human species may be incorporated.
[0252] The term "incorporation" of CDR's or hypervariable regions
as used herein encompasses any means by which the non-human CDR's
are situated with the human acceptor framework. It will be
appreciated that this can be achieved in various ways, for example,
nucleic acids encoding the desired amino acid sequence can be
generated by mutating nucleic acids encoding the non-human variable
domain sequence so that the framework residues thereof are changed
to human acceptor framework residues, or by mutating nucleic acid
encoding the human variable domain sequence so that the CDR's are
changed to non-human residues, or by synthesizing nucleic acids
encoding the desired sequence. In one embodiment the final sequence
is generated in silico.
[0253] The present invention is now described by way of example
only. The appended claims may include a generalisation of one of
more of the following examples.
EXAMPLES
Example 1
Monoclonal Antibody Generation and Selection
1.1 Immunisation Strategies
[0254] The anti human BCMA mAb murine parental CA8 was identified
from hybridomas derived from mice immunized with full length human
BCMA. A BALB/c mouse was immunized i.p. with 25 .mu.g of
recombinant (rBCMA) protein combined with CFA. The mouse was
boosted three times at one-month intervals with 25 .mu.g of full
length rBCMA protein+10 .mu.g monophosphoryl lipid A-stable
emulsion (MPL-SE) (Corixa Corporation, Seattle, Wash.) and given a
pre-fusion boost of 30 .mu.g rBCMA protein i.v. 3 days prior to
fusion. Hybridomas were either generated and cloned using the
ClonaCell-HY hybridoma cloning kit (StemCell Technologies,
Vancouver, BC) or using a conventional method. In the conventional
method, B cells from the spleens of the immunized animals were
fused with Sp2/0 myeloma cells in the presence of PEG
(Sigma-Aldrich, St. Louis, Mo.). After overnight recovery, fused
cells were plated at limiting dilution in 96-well plates and
subjected to hypoxanthine-aminopterin-thymidine selection.
Hybridoma culture supernatants were examined for the presence of
anti-BCMA antibodies by ELISA and flow cytometry.
[0255] The anti human BCMA mAb murine parental S307118G03 was
identified from hybridomas derived from SJL mice immunized with
recombinant human BCMA/TNFRSF17-Fc chimera (R&D 193-Fc) using
the RIMMS method (Rapid immunisation multiple sites). At Day 0, 5
ug protein per mouse was emulsified in AS02a adjuvant at 2 sites on
back (over haunches and over shoulders) and subjacent to the major
lymph nodes at 4 sites on front. On day 6 and day 11 2.5 ug protein
per mouse in RIBI adjuvant was injected subjacent to the major
lymph nodes at 4 sites on front. On day 14 the animals were
sacrificed. The lymph nodes and spleen were excised, disrupted and
a PEG1500 induced somatic cell fusion performed using a 3:1 ratio
with mouse myeloma cells X63 AG8 653.GFP.Bcl-2.11 (BioCat 112754;
R17209/58). The fusion was plated out into 10.times.96 well plates
and screened directly from these.
[0256] The anti human BCMA mAb murine parental S336105A07 was
identified from hybridomas derived from identical immunisations.
The lymph nodes and spleen were excised at day 14, disrupted, and a
Cytopulse electrofusion was performed using a 1:1 ratio with mouse
myeloma cells X63 AG8 653.GFP.Bcl-2.11 (BioCat 112754; R17209/58).
The fusion was plated out into omnitrays containing semi solid
medium prior to picking into 10.times.96 well plates and was
screened directly from these 5 days later.
[0257] The anti human BCMA murine parental mAbs S332121F02 and
S332126E04 were identified from hybridomas derived from SJL mice
immunized with recombinant Fc fusion of the extracellular domain of
human BCMA (4-53) BCMA using the RIMMS method (Rapid immunisation).
At Day 0, 5 ug protein per mouse was emulsified in AS02a adjuvant
at 2 sites on back (over haunches and over shoulders) and subjacent
to the major lymph nodes at 4 sites on front. On day 6 5 ug
recombinant cyno BCMA-Fc protein per mouse in RIBI adjuvant was
injected subjacent to the major lymph nodes at 4 sites on front. On
day 11 2.5 ug recombinant human BCMA-Fc and 2.5 ug recombinant cyno
BCMA-Fc per mouse in RIBI adjuvant was injected subjacent to the
major lymph nodes at 4 sites on front. On day 14 the animals were
sacrificed and cells treated as for S307118G03.
[0258] The anti human BCMA murine parental mAb S322110D07 was
identified from hybridomas derived from SJL mice immunised with
recombinant Fc fusion of the extracellular domain of human BCMA
(4-53) in complex with recombinant human April (R&D 5860-AP/CF)
premixed at 1:1 molar ratio. The mice were immunized i.p. with 5 ug
April/Cyno BCMA-Fc complex in PBS, suspended in RIBI adjuvant, 100
ul dose per mouse and boosted 3 times at 3-4 week intervals with
2.5 ug April/Cyno BCMA-Fc complex in PBS, suspended in RIBI
adjuvant, 100 ul dose per mouse injected via intraperitoneal route
and given a pre-fusion boost of the same immunogen 1 day prior to
fusion and treated as for S307118G03.
[0259] The anti human BCMA mAb murine parental S335115G01 and
S335122F05 were identified from hybridomas derived from SJL mice
immunized with a mixture of recombinant Fc fusion of the
extracellular domain of human BCMA (4-53) and recombinant Fc fusion
of the extracellular domain of cyno BCMA (4-52) using the RIMMS
method (Rapid immunisation multiple sites). At Day 0, 2, 5 ug of
each protein per mouse was emulsified in AS02a adjuvant and
injected at 2 sites on the back (over haunches and over shoulders)
and subjacent to the major lymph nodes at 4 sites on front. On day
6 and day 11 2.5 ug of each protein per mouse in RIBI adjuvant was
injected subjacent to the major lymph nodes at 4 sites on front. On
day 14 the animals were sacrificed. The lymph nodes and spleen were
excised, disrupted and a Cytopulse electrofusion was performed
using a 1:1 ratio with mouse myeloma cells X63 AG8 653.GFP.Bcl-2.11
(BioCat 112754; R17209/58). The fusion was plated out into
omnitrays containing semi solid medium prior to picking into
32.times.96 well plates and was screened directly from these 5 days
later.
Example 2
Humanisation
2.1 Cloning of CA8 Hybridoma Variable Regions
[0260] Total RNA was extracted from CA8 hybridoma cells, heavy and
light variable domain cDNA sequence was then generated by reverse
transcription and polymerase chain reaction (RT-PCR). The forward
primer for RT-PCR was a mixture of degenerate primers specific for
murine immunoglobulin gene leader-sequences and the reverse primer
was specific for the antibody constant regions. Reverse primers
specific for IgG1, IgG2a and IgG2b were used in this case as the
isotype was unknown. To design the primers, DNA multiple sequence
alignments of the leader sequences of the mouse V.sub.H and V.sub.k
genes were generated.
2.2 Cloning of Chimeric CA8
[0261] The DNA expression constructs encoding the chimeric antibody
were prepared de novo by build-up of overlapping oligonucleotides
including restriction sites for cloning into mammalian expression
vectors as well as a human signal sequence. HindIII and SpeI
restriction sites were introduced to frame the VH domain containing
the signal sequence for cloning into mammalian expression vectors
containing the human .gamma.1 constant region. HindIII and BsiWI
restriction sites were introduced to frame the VL domain containing
the signal sequence for cloning into mammalian expression vector
containing the human kappa constant region.
2.3 Cloning of the Humanised CA8 Variants
[0262] The DNA expression constructs encoding the humanised
antibody variants were prepared de novo by build-up of overlapping
oligonucleotides including restriction sites for cloning into
mammalian expression vectors as well as a human signal sequence.
HindIII and SpeI restriction sites were introduced to frame the VH
domain containing the signal sequence for cloning into mammalian
expression vectors containing the human .gamma.1 constant region.
HindIII and BsiWI restriction sites were introduced to frame the VL
domain containing the signal sequence for cloning into mammalian
expression vector containing the human kappa constant region.
2.4 Expression of the Recombinant CA8 Antibodies (Including
Antibody Quantification)
[0263] Expression plasmids encoding the heavy and light chains
respectively were transiently co-transfected into HEK 293 6E cells
and expressed at small scale to produce antibody. Antibodies were
quantified by ELISA. ELISA plates were coated with anti human IgG
(Sigma 13382) at 1 mg/ml and blocked with blocking solution (4% BSA
in Tris buffered saline). Various dilutions of the tissue culture
supernatants were added and the plate was incubated for 1 hour at
room temperature. Dilutions of a known standard antibody were also
added to the plate. The plate was washed in TBST and binding was
detected by the addition of a peroxidise labelled anti human kappa
light chain antibody (Sigma A7164) at a dilution of 1/1000 in
blocking solution. The plate was incubated for 1 hour at room temp
before washing in TBST. The plate was developed by addition of OPD
substrate (Sigma P9187) and colour development stopped by addition
of 2M H2SO4. Absorbance was measured at 490 nm and a standard curve
plotted using data for the known standard dilutions. The standard
curve was used to estimate the concentration of antibody in the
tissue culture supernatants. Larger scale antibody preparations
were purified using protein A and concentrations were measured
using a Nanodrop (Thermo Scientific).
TABLE-US-00002 TABLE 1 Design of CA8 variable heavy and light
humanised variants Humanised Backmutations VH Template (Kabat#) J0
Straight graft of CA8 VH CDRs None onto IGHV1_69 + JH1 minigene J1
J0 G27Y, S30T J2 J1 A93T J3 J2 A24G, K73T J4 J3 M48I, V67A, J5 J3
N99D J6 J0 N99D J7 J1 N99D J8 J2 N99D J9 J4 N99D M0 Straight graft
of CA8 VL CDRs None onto IGKV1_39 + JK2 minigene M1 M0 F71Y M2 M1
M4L, K45E,
2.5 Defucosylated Antibody Production
[0264] To generate defucosylated antibodies the heavy and light
chains respectively were co-transfected into CHO DG44 MS705 BioWa
cells and expressed at scale to produce antibody. Briefly, 30 .mu.g
DNA was linearised overnight with Not1, the DNA was ethanol
precipitated and re-dissolved in TE buffer. From culture,
2.4.times.107 BioWa DG44 cells were obtained and washed in 14 ml of
warmed PBS-sucrose. The cells were spun and the pellet resuspended
in 1.6 ml of PBS-sucrose. Half (0.8 ml) of aforementioned cells,
suspended in PBS-sucrose, were added to a BioRad cuvette with the
30 .mu.g of linearised DNA (in 50 .mu.l TE buffer). A BioRad
GenePulser was programmed to 380V with a capacitance of 25 .mu.F
and the cuvette was entered for electroporation. The resulting 850
ul of electroporated cells and DNA were added to (80 ml) warmed
SFM512 medium (including phenol red, 2.times.HT (nucleosides),
glutamax and Gibco supplement4). Finally, the resulting 80 ml of
cell suspension was transferred.left brkt-bot. (150 .mu.l/well) to
each well of one of 4.times.96-well plates. After 48 hours, the
medium was changed to nucleoside free by removing approximately 130
.mu.l of conditioned and replacing with 150 .mu.l of fresh
selection medium SFM512 medium (including phenol red and glutamax).
Every 3-4 days, 130-150.mu.l of conditioned medium was removed and
replaced with fresh, selection medium. Wells were monitored for
colour change and assayed for IgG concentration as discussed
previously.
2.6 Additional Antibodies--Cloning of Hybridoma Variable
Regions
[0265] Total RNA was extracted from S307118G03, S332121F02,
S332126E04, S322110D07, S336105A07, S335115G01 and S335122F05
hybridoma cells. Heavy and light variable domain cDNA sequence was
then generated by reverse transcription and polymerase chain
reaction (RT-PCR).
[0266] The forward primer for RT-PCR was a mixture of degenerate
primers specific for murine immunoglobulin gene leader-sequences
and the reverse primer was specific for the antibody constant
regions, in this case isotype IgG2a. Primers were designed based on
a strategy described by Jones and Bendig (Bio/Technology 9:88,
1991). RT-PCR was carried out for both V-region sequences to enable
subsequent verification of the correct V-region sequences. DNA
sequence data was obtained for the V-region products generated by
the RT-PCR.
2.7 Additional Antibodies--Cloning of the Chimeras
[0267] The DNA expression constructs encoding the chimeric
antibodies were prepared de novo by infusion advantage PCR cloning
(Clonetech) of the V-gene PCR products into mammalian expression
vectors. This cloning method enabled fusion the murine variable
regions to human IgG1 H chain and kappa L chain constant
regions.
2.8 S307118G03--Cloning of the Humanized Variants
[0268] Cloning was carried out as for paragraph 2.3.
2.9 S307118G03 Expression of the Recombinant Antibodies
[0269] Expression plasmids encoding the relevant heavy and light
chains (listed in Table 8 below) were transiently co-transfected
into HEK 293 6E cells and expressed at small scale to produce
antibody. The antibodies were Protein A purified from the
supernatants and quantified using the Nanodrop
spectrophotometer.
[0270] 8 below) were transiently co-transfected into HEK 293 6E
cells and expressed at small scale to produce antibody. The
antibodies were Protein A purified from the supernatants and
quantified using the Nanodrop spectrophotometer.
Example 3
Conjugation of Antibodies to vcMMAE and mcMMAF to Form Antibody
Drug Conjugates (ADC)
TABLE-US-00003 [0271] TABLE B Chemical structures of drug-linkers
##STR00003## ##STR00004##
[0272] Gammabind Plus Protein G Sepharose (GE Healthcare) resin
slurry (75 uL) was added to a each well of a deep well (2 mL
capacity) filter plate. The antibodies to be conjugated were
grouped by species and isotype and up to 0.5 mg of each antibody
transferred to each well of the plate. Each antibody was
transferred to two separate wells to facilitate the preparation of
two conjugates, with the drug-linkers SGD-1006 and SGD-1269. The
filter plate was then shaken at 1200 RPM for 2 hours at 5.degree.
C. to bind the antibodies to the resin. The filter plate was then
centrifuged at 500.times.g for 3 minutes to ensure complete
pulldown of all fluids and resin to the bottom of the each
well.
[0273] The bound antibodies were then reduced by adding 500 uL of
10 mM TCEP in 100 mM KPO4, 150 mM NaCl, pH 7, 1 mM EDTA and shaking
for 30 minutes at 22.degree. C. Following reduction, the plate was
again centrifuged to remove the TCEP solution and subsequently
washed with PBS+1 mM EDTA, 1 mL per well. The wash solution was
removed by centrifugation and the process repeated 3 times for a
total of 4 washes. The bound and reduced antibodies were then
conjugated using a mixture of NEM and drug linker prepared in
accordance with the mole fractions indicated in Table 2.
TABLE-US-00004 TABLE 2 Antibody Reducible SGD-1006 mole SGD-1269
mole (species/isotype) Disulfides fraction fraction Human IgG1* 4
0.675 0.688 Murine IgG1 5 0.500 0.586 Murine IgG2a 5 0.500 0.586
Murine IgG2b 6 0.463 0.481 *also for murine/human IgG1
chimerics
[0274] Separate mixtures of NEM and drug linker were thus prepared
for each antibody species/isotype using 10 mM DMSO stock solutions
of SGD-1006, SGD-1269 (See Table B) and NEM. When mixed at the
appropriate ratio the total maleimide concentration was therefore
still 10 mM, and this value was used to calculate the volume of
maleimide solution to be added to each well. For example for a
murine IgG1 with 5 reducible disulfides (10 available thiols when
reduced) 0.5 mg of antibody at 150 kD is 3.33 nmol corresponding to
33.3 nmol of thiol. A 3-fold excess is therefore 100 nmol of total
maleimide or 10 .mu.L of the 10 mM drug linker/NEM mix. For the
SGD-1269 conjugate this mix would then be prepared with 5.86 .mu.L
of SGD-1269 and 4.14 .mu.L of NEM. The maleimide mix would then be
diluted into 500 .mu.L of PBS prior to addition to the immobilized
reduced antibody. In practice, since multiple antibodies of each
isotype were conjugated simultaneously a single SGD-1269/NEM mixed
solution for each isotype was prepared by multiplying the number of
wells containing that isotype by 10 .mu.L per well then diluting
into a volume of PBS equal to 500 .mu.L times the number of wells.
In like fashion a total of eight drug-linker/NEM mixes were
prepared--four with SGD-1006 and four with SGD-1269--and diluted
into PBS. These mixes were then added to the reduced antibodies
(500 .mu.L per well) and the plate was shaken for 30 minutes at
22.degree. C. The plate was then centrifuged as above to remove the
excess reaction solution, and subsequently washed 4 times with PBS
as before. The bound ADCs were then eluted by adding 200 uL of 50
mM glycine pH 2.5 to each well and shaking the plate for 3 minutes
at 1200 RPM. While shaking 20 uL of neutralization buffer (1M
potassium phosphate, pH 7.4, 500 mM NaCl, 0.2% Tween-20) was added
to each well of a 1 mL collection plate. The ADCs were then eluted
into the collection plate by spinning at 1500.times.g for 6
minutes. The collection plate was then shaken briefly to ensure
complete mixing of the neutralization buffer.
[0275] The concentration of each ADC was then determined with an
absorbance plate reader by transferring the solutions into a UV
assay plate (Costar model 3635, Corning) and measuring the optical
density at 280 nm. An average IgG extinction coefficient of 1.45 mL
mg-1 cm-1 was used to provide an adequate estimation of ADC
concentration across the panel. To confirm successful conjugation,
a reversed phase protein HPLC method (described below) was used to
estimate the drug loading of the isotype controls. For the plate
containing the humanization variants of CA8 this method was used to
estimate the loading of all ADCs directly.
[0276] The reversed phase protein chromatography method for
determining drug loading employs the PLRP-S polymeric stationary
phase (Agilent Technologies). Since the antibodies were fully
reduced during the conjugation process all of the antibody subunits
elute from the column as single polypeptide chains allowing the
subpopulations of light and heavy chain species with varying levels
of drug loading to be evaluated separately. Thus, the analysis of
these data allow for the calculation of the average light chain
drug loading and the average heavy chain drug loading as
independent factors which can then be combined to determine average
antibody drug loading with the basic knowledge that each antibody
is comprised of two light and two heavy chains. The chromatographic
conditions were as follows: A PRLP-S column, 1000 .ANG.,
50.times.2.1 mm, 8 um particle size (Agilent Technologies) with
water+0.05% TFA as mobile phase A and acetonitrile+0.01% TFA as
mobile phase B; elution with a linear gradient of 27% B to 42% B in
12.5 minutes.
[0277] Anti-BCMA antibodies were conjugated with SGD-1006 and
SGD-1269 in three separate batches over a period of seven months.
In the first batch a total of 29 antibodies were conjugated
(resulting in 58 ADCs). The drug loading of each isotype control
determined by PLRP chromatography and the data are summarized in
Table 3.
TABLE-US-00005 TABLE 3 Isotype SGD-1006 loading SGD-1269 loading
cIgG1 (control P) 4.23 4.35 cIgG1 (control M) 4.42 4.41 mIgG1 4.26
4.04 mIgG2a 4.51 4.57 mIgG2b 4.39 4.18
[0278] For the second batch an additional 25 antibodies were
conjugated (resulting in 50 ADCs). The drug loading of each isotype
control was again determined by PLRP chromatography and the data
are summarized in Table 4.
TABLE-US-00006 TABLE 4 SGD-1269 Isotype SGD-1006 loading loading
cIgG1 3.96 3.78 mIgG1 3.95 3.32 mIgG2a 4.53 3.60 mIgG2b 4.32
3.49
[0279] In the third batch 30 antibodies were conjugated (resulting
in 60 ADCs), including 13 humanized variants of CA8. In this final
batch, the drug loading of all ADCs were determined and are
summarized in the following two plate maps. (Table 5 & 6)
TABLE-US-00007 TABLE 5 drug loading 1 2 3 4 5 6 7 8 9 10 A 3.7 4.0
3.6 3.8 3.8 3.5 3.9 2.8 3.8 3.8 B 3.7 3.6 3.5 3.7 4.0 3.4 3.7 3.3
3.8 3.9 C 3.6 3.8 3.5 3.7 3.6 3.3 3.8 4.7 3.8 3.7 D 3.4 3.6 3.6 3.9
3.9 3.4 3.2 4.8 3.8 3.9 E 3.9 3.8 3.9 3.4 3.6 3.3 3.7 3.4 F 3.7 4.0
3.6 3.5 3.8 3.7 G 3.6 3.6 3.4 3.7 H 3.6 3.6 SGD-1006 (vc-MMAE) ADCs
SGD-1269 (mc-MMAF) ADCs 3.7 3.8 3.6 3.8 3.4 3.7 3.8 3.7 4.1% 5.1%
3.4% 4.8% 2.8% 8.5% 24.1% 3.4%
TABLE-US-00008 TABLE 6 mIgG1 mIgG2a mIgG2b humanized mIgG1 mIgG2a
mIgG2b humanized 1 2 3 4 5 6 7 8 9 10 A control control control CA8
J6M0 CA8 J8M2 control control control CA8 J6M0 CA8 J8M2 B
S336106D07 S336105A07 S336107G08 CA8 J6M1 CA8 J9M0 S336106D07
S336105A07 S336107G08 CA8 J6M1 CA8 J9M0 C S335115G03 S335122F05
S336104A09 CA8 J6M2 CA8 J9M1 S335115G03 S335122F05 S336104A09 CA8
J6M2 CA8 J9M1 D S335115G01 S335128A12 S335107H11 CA8 J7M0 CA8 J9M2
S335115G01 S335128A12 S335107H11 CA8 J7M0 CA8 J9M2 E S335106E08
S335119E11 CA8 J7M1 CA8 Fc ENH S335106E08 S335119E11 CA8 J7M1 CA8
Fc ENH F S335132E01 CA8 J7M2 GRITS28785 S335132E01 CA8 J7M2
GRITS28785 G S341106G02 CA8 J8M0 S341106G02 CA8 J8M0 H CA8 J8M1 CA8
J8M1 SGD-1006 (vc-MMAE) ADCs SGD-1269 (mc-MMAF) ADCs
[0280] Mean drug loading and % CV are indicated for each isotype
series at the bottom. An uncharacteristically large variability in
drug loading was observed for the SGD-1269 ADCs prepared with
mlgG2b antibodies; the reason for this is unclear. Also, the
Fc-enhanced CA8 antibodies yielded somewhat lower drug loading
levels than the other CA8 human variants; to address this,
additional Fc-enhanced CA8 was conjugated in a solution-phase
reaction to better match the drug loading achieved for the other
antibodies.
Example 4
Binding Data
4.1 FMAT Binding Assay to Show Binding of Chimeric CA8 to Cells
Expressing Human or Cyno BCMA.
[0281] Cryopreserved transfected human, cyno BCMA and mock
transfected HEK293 cells were recovered from LN2 storage. Assay
wells were prepared with human chimeric CA8 antibody, at a range of
different concentrations, mixed with human BCMA HEK293, cyno BCMA
HEK293 and mock transfected cells respectively. Anti-human IgG FMAT
Blue secondary conjugate was added for detection of human chimeric
CA8. The assay plates were left for a minimum of 90 minutes before
the result was read on the AB18200 (FMAT) plate reader.
[0282] This showed that the CA8 antibody in chimeric form binds
well to both human and cyno BCMA proteins expressed on HEK293
cells.
[0283] Results are shown in FIG. 1.
4.2 ELISA Experiment Showing Binding of Chimeric CA8 to Recombinant
BCMA Protein
[0284] Chimeric CA8 antibodies were tested for binding to human
BCMA and cyno BCMA expressed as Fc fusions. Human BCMA-Fc and cyno
BCMA-Fc were coated to ELISA plates and the plates were blocked
using BSA to reduce non specific binding. CA8 chimeric antibodies
were added in a concentration range from 5 ug/ml to 0.1 ug/ml to
the human and cyno BCMA coated ELISA plates. Any bound human
chimeric CA8 antibody was detected using anti-human IgG HRP
conjugated secondary antibody as appropriate. HRP substrate (TMB)
was added to develop the ELISA. This showed that CA8 antibody binds
to recombinant human and cyno BCMA in an ELISA assay.
[0285] Results are shown in FIG. 2.
4.3 Biacore Experiment to Show CA8 Antibody Binding to BCMA and
TACI Proteins to Determine Cross Reactivity with TACI Protein.
[0286] CA8 chimera antibody was injected and captured on protein A.
(A protein A derivitised sensorchip was used). Residual protein A
binding was blocked with an injection of a high concentration of
human IgG solution. BCMA-Fc, TACI-Fc or BAFF-R-Fc solutions were
then tested for binding to the antibody. The 3 proteins were
injected in sequence and binding events were measured. The surface
was regenerated between injection of each protein.
[0287] Sensorgrams were analysed in the Biaevaluation program.
Double reference subtraction was done to remove instrument noise
and any non-specific binding from the sensorgram curves.
[0288] This showed that CA8 was specific for binding to BCMA
binding and not to TACI and BAFFR.
[0289] Binding of the CA8 antibody to BCMA-Fc, TACI-Fc and
BAFF-R-Fc was plotted out as shown in FIG. 3.
4.4 Cell Binding and Neutralisation Data
4.4.1 Binding of Murine Anti BCMA Antibodies to Multiple Myeloma
Cells and BCMA Expressing Cells
[0290] Multiple myeloma cell line H929 and ARH77-hBCMA 10B5 BCMA
expressing transfectant cells were stained with murine S332211 D07,
S3332121F02 or S332126E04 or murine isotype control at 5 .mu.g/mL.
Multiple myeloma cell line H929 was stained with murine S307118G03.
Cells were incubated for 20 mins at room temperature (RD and then
washed with FACS buffer (PBS+0.5% BSA+0.1% sodium azide) to remove
unbound antibody. Cells were incubated with a secondary PE labelled
anti-mouse IgG antibody for 15 minutes at RT and then washed with
FACS buffer to remove unbound antibody. Cells were analysed by FACS
to detect antibody bound to the cells.
[0291] The results (FIG. 4) showed that all 4 murine antibodies
bound to the H929 multiple myeloma cell line and the three
antibodies tested on ARH77 BCMA transfected cells bound to
these.
4.4.2 Binding Curve of Chimeric CA8 to Multiple Myeloma Cells as
Determined by FACS
[0292] A panel of multiple myeloma cell lines were used to
determine the binding of chimeric CA8. Cell lines H929, OPM-2,
JJN-3 and U266 were stained with either chimeric CA8 or irrelevant
antibody (Synagis) at varying concentrations for 20 minutes at RT.
Cells were then washed with FACS buffer (PBS+0.5% BSA+0.1% sodium
azide) to remove unbound antibody. Cells were incubated with a
secondary PE labelled anti-human IgG antibody for 15 minutes at RT
and then washed with FACS buffer to remove unbound antibody. Cells
were analysed by FACS and mean fluorescence intensity (MFI) values
measured to determine binding.
[0293] Results showed that chimeric CA8 bound to multiple myeloma
cell lines H929, OPM-2, JJN-3 & U266 in a dose dependent manner
(FIG. 5).
4.4.3 Binding of Humanised CA8 to BCMA Transfected Cells as
Determined by FACS
[0294] ARH77-hBCMA 10B5 BCMA expressing transfectant cells or H929
cells were stained with either chimeric CA8 or humanised variants
of CA8 designated J6M0, J6M1, J6M2, J9M0, J9M1, J9M2 at varying
concentrations for 20 minutes at RT. Cells were then washed with
FACS buffer (PBS+0.5% BSA+0.1% sodium azide) to remove unbound
antibody. Cells were incubated with a secondary PE labelled
anti-human IgG antibody for 15 minutes at RT and then washed with
FACS buffer to remove unbound antibody. Cells were analysed by FACS
and mean fluorescence intensity (MFI) values measured to determine
binding.
[0295] Results showed that chimeric CA8 and all antibodies tested
apart from J9M2 bound to ARH77-hBCMA 10B5 BCMA expressing
transfectant cells and H929 cells in a dose dependent manner (FIG.
6).
4.5 Demonstration of Ability of CA8 and the Humanised Version J6M0
to Neutralise Binding of BAFF or APRIL to Recombinant BCMA.
[0296] The aim of this assay was to assess the ability of antibody
CA8, and humanised version J6M0 in both wild type and afucosylated
(Potelligent) form, at various concentrations, to neutralise the
binding ability of either BCMA ligand, BAFF or APRIL.
[0297] 96 well flat bottomed plates were coated overnight with 1
.mu.g/mL solution of recombinant human BCMA Fc 4-53 in PBS.
Following a wash step using 0.05% TWEEN20, plates were blocked with
2% Bovine Serum Albumin solution in PBS for 1 hour at room
temperature. Plates were washed as before and 40 .mu.L of each
antibody (murine IgG, murine CA8, and chimeric CA8), starting at 10
.mu.g/mL, titrated at 1 in 2 in duplicate was added to the relevant
wells and incubated for 1 hour at room temperature. 40 .mu.L of 2%
BSA was added to the relevant control wells. 10 .mu.L of either
recombinant human BAFF (2149-BF/CF, R&D Systems) or recombinant
human APRIL (5860-AP/CF, R&D Systems) was added at 30 ng/mL and
750 ng/mL respectively, giving a final concentration of 6 ng/mL and
150 ng/mL respectively in each well. Equivalent volume of 2% BSA
was added to the relevant control wells. Plates were allowed to
incubate for 2 hours at room temperature, after which they were
washed as before. Biotinylated anti-human ligand (BAFF BAF124 or
APRIL BAF884, R&D Systems) was added to the relevant wells at
50 ng/mL and incubated for 1 hour. Following a wash step, 50 .mu.L
of a 1:4000 dilution of Streptavidin-HRP (Amersham RPN4401) was
added to each well and incubated for 30 minutes at room
temperature. The wash process was repeated again followed by the
addition of 100 .mu.L of Tetramethylbenzidine substrate solution
(T8665, Sigma) into each well. Plates were incubated for 20-25
minutes at room temperature, wrapped in foil. The reaction was
stopped with the addition of 100 .mu.L of 1M H.sub.2SO.sub.4.
Optical density was determined at 450 nm using Spectromax reader.
See FIGS. 7A and B.
[0298] In a plate based assay for neutralisation of binding of BAFF
or APRIL to BCMA, the EC50 values calculated for chimeric CA8 were
0.695 .mu.g/mL and 0.773 .mu.g/mL respectively. The values for the
humanised J6M0 were 0.776 ng/ml and 0.630 ng/ml. The values for the
J6M0 potelligent version were 0.748 and 0.616 ng/ml
respectively.
4.6 Effect of Chimerised CA8 and Humanised J6M0 BCMA Antibody on
BAFF or APRIL Induced Phosphorylation of NFkB in H929 Cells.
[0299] In one set of experiments, H-929 cells were plated at 75,000
cells/well in a 96 well plate in serum free medium. The chimeric
CA8 antibody was added 24 hours later to give final well
concentrations up to 200 ug/ml. Ten minutes later, BAFF or APRIL
ligand were added to the cells to give final well concentrations of
0.6 or 0.3 ug/ml respectively. After 30 minutes the cells were
lysed and phosphorylated NfkappaB levels measured using a MSD
pNFkappaB assay.
[0300] The chimeric BCMA antibody CA8 neutralised both BAFF and
APRIL induced NfkappaB cell signalling in H-929 cells. It was
particularly potent at neutralising BAFF induced NfkappaB cell
signalling in this cell type with a mean IC50 of 10 nM, compared to
257 nM for APRIL induced NfkappaB cell signalling.
Meaned Data for 2 Experiments
[0301] IC50s were 10 nM for BAFF induced NfkappaB neutralisation
and 257 nM for APRIL induced NfkappaB neutralisation (mean of 2
independent experiments) are shown in Table 7.
TABLE-US-00009 TABLE 7 BAFF induced IC50 APRIL induced IC50 ug/ml
nM ug/ml nM BCMA antibody CA8 1.5 10 38.5 256.7
[0302] A further set of experiments were carried out to aim to
understand why there was such a discrepancy between the potency in
neutralisation of APRIL and BAFF in the cell based system.
Following the discovery of the soluble form of BCMA the
experimental design was changed to include a step where the H929
cells were washed prior to the assay to reduce the interference
from the antibody binding to soluble BCMA. H-929 cells were washed
3 times to remove any sBCMA and resuspended in serum free medium.
J6M0 potelligent antibody was added to a 96 well plate to give a
final well concentrations up to 100 ug/ml along with BAFF or APRIL
ligand to give a final well concentration of 0.6 or 0.2 ug/ml
respectively. H-929 cells were then plated at 7.5.times.104
cells/well in serum free medium. 30 minutes later the cells were
lysed and phosphorylated NFkappaB levels measured using a MSD
pNFkappaB assay. This is data from one experiment. Each data point
is the mean/sd of two replicates. The data from this experiment is
shown in FIG. 7c. The IC50s for inhibition of BAFF and APRIL
signalling were determined as 0.91 ug/ml and 2.43 ug/ml
respectively.
4.7 ProteOn Analysis of Anti-BCMA CA8 Chimeric and Humanised
Constructs
[0303] The initial screen of CA8 chimeric and humanised variants
was carried out on the ProteOn XPR36 (Biorad). The method was as
follows; Protein A was immobilised on a GLC chip (Biorad, Cat No:
176-5011) by primary amine coupling, CA8 variants were then
captured on this surface and recombinant human BCMA (in house or
commercial US Biological, B0410) materials (run 2 only)) passed
over at 256, 64, 16, 4, 1 nM with a 0 nM injection (i.e. buffer
alone) used to double reference the binding curves, the buffer used
is the HBS-EP buffer. 50 mM NaOH was used to regenerate the capture
surface. The data was fitted to the 1:1 model using the analysis
software inherent to the ProteOn XPR36. Run 1 corresponds to the
first screen of humanised CA8 variants (JO to J5 series) and run 2
to the second screen of humanised CA8 variants (J5 to J9 series).
Both runs were carried out at 25.degree. C.
[0304] The data obtained from run1 are set out in Table 8 and data
from run 2 are set in Table 9 Several molecules in the Run 2 (Table
09) failed to give affinity values measurable by ProteOn, this was
due to the off-rate being beyond the sensitivity of the machine in
this assay, this does however indicate that all these molecules
bind tightly to recombinant human BCMA. From Run 1 the data
indicates that some constructs did not show any binding to
recombinant cyno BCMA.
TABLE-US-00010 TABLE 8 Run 1-Kinetics analyses of anti-BCMA
molecules against Recombinant Human BCMA Human in house BCMA Cyno
in house BCMA KD KD Sample name ka kd (nM) ka kd (nM) CA8 humanised
J5M0 2.16E+05 1.88E-05 0.087 3.25E+05 8.14E-06 0.025 CA8 humanised
J5M2 2.67E+05 3.21E-05 0.12 4.30E+05 4.70E-05 0.109 CA8 humanised
J5M1 2.97E+05 4.32E-05 0.145 4.81E+05 5.41E-05 0.112 CA8 humanised
J4M1 2.54E+05 7.04E-05 0.278 3.50E+05 7.10E-05 0.203 CA8 humanised
J4M2 2.51E+05 7.06E-05 0.281 3.44E+05 6.15E-05 0.179 CA8 humanised
J0M2 2.25E+05 6.97E-05 0.31 3.26E+05 1.84E-04 0.563 CA8 humanised
J3M2 2.66E+05 9.64E-05 0.362 3.69E+05 5.87E-05 0.159 CA8 humanised
J0M1 2.31E+05 8.60E-05 0.373 3.32E+05 1.67E-04 0.503 CA8 humanised
J0M0 2.45E+05 1.06E-04 0.435 3.58E+05 2.32E-04 0.648 CA8 humanised
J3M1 2.85E+05 1.25E-04 0.438 4.04E+05 7.93E-05 0.196 CA8 humanised
J2M2 2.05E+05 9.87E-05 0.482 2.98E+05 3.17E-05 0.106 CA8 Chimera
2.41E+05 1.25E-04 0.519 3.82E+05 1.74E-04 0.457 CA8 humanised J2M1
2.04E+05 1.72E-04 0.842 2.96E+05 6.46E-05 0.218 CA8 humanised J4M0
2.42E+05 2.20E-04 0.906 3.34E+05 2.89E-04 0.866 CA8 humanised J1M2
2.15E+05 2.46E-04 1.14 3.19E+05 9.67E-05 0.303 CA8 humanised J3M0
2.08E+05 2.85E-04 1.37 2.93E+05 1.54E-04 0.526 CA8 humanised J1M1
2.27E+05 3.43E-04 1.51 3.33E+05 1.47E-04 0.442 CA8 humanised J2M0
1.95E+05 3.77E-04 1.94 2.81E+05 1.51E-04 0.538 CA8 humanised J1M0
1.78E+05 5.02E-04 2.82 2.47E+05 2.10E-04 0.849 S307118G03 Chimera
4.75E+05 1.95E-03 4.11 No Analysable Binding S307118G03 humanised
H3L1 4.69E+05 2.28E-03 4.86 No Analysable Binding S307118G03
humanised H3L0 2.86E+05 1.52E-03 5.31 No Analysable Binding
S307118G03 humanised H2L0 3.78E+05 2.41E-03 6.36 No Analysable
Binding S307118G03 humanised H2L1 3.38E+05 2.15E-03 6.37 No
Analysable Binding S307118G03 humanised H4L1 No Analysable Binding
No Analysable Binding
TABLE-US-00011 TABLE 9 Run 2-Kinetics analyses of anti-BCMA
molecules against Recombinant Human BCMA commercial human Human in
house BCMA BCMA KD KD Cyno in house BCMA Sample Name ka kd (nM) ka
kd (nM) ka kd KD (nM) CA8 Chimera 2.51E+05 1.03E-04 0.412 7.05E+05
9.79E-05 0.139 5.89E+04 1.21E-04 2.060 CA8 humanised J6M1 2.17E+05
2.70E-05 0.124 5.92E+05 3.75E-05 0.063 4.88E+04 2.58E-04 5.300 CA8
humanised J6M0 2.40E+05 7.40E-05 0.308 6.23E+05 5.37E-05 0.086
5.64E+04 3.18E-04 5.630 CA8 humanised J6M2 2.01E+05 4.06E-05 0.202
5.63E+05 3.97E-05 0.071 4.41E+04 3.02E-04 6.860 S307118G03 H5L0 No
Analysable Binding V weak signal No Analysable Binding S307118G03
H5L1 No Analysable Binding V weak signal No Analysable Binding
S307118G03Chimera 4.79E+05 1.65E-03 3.44 1.55E+06 1.48E-03 0.956 No
Analysable Binding
[0305] For antibodies J8M0, J9M0, J8M1, J9M2, J7M2, J5M0, J7M1,
J7M0, J8M2, J9M1, J5M2, J5M1 the off rate was beyond the
sensitivity of the assay hence no data shown.
4.8 BIAcore Analysis of Anti-BCMA CA8 Chimeric and Humanised
Constructs (J7 to J9 Series)
[0306] Protein A was immobilised on a CM5 chip (GE Healthcare, Cat
No: BR-1005-30) by primary amine coupling and this surface was then
used to capture the antibody molecules. Recombinant human BCMA (US
Biological, B0410) was used as analyte at 256 nM, 64 nM, 16 nM, 4
nM and 1 nM. Regeneration of the capture surface was carried out
using 50 mM NaOH. All binding curves were double referenced with a
buffer injection (i.e. 0 nM) and the data was fitted to the using
the 1:1 model inherent to T100 evaluation software. The run was
carried out at 37.degree. C., using HBS-EP as the running
buffer.
[0307] The results showed the molecules tested with the exception
of J9M2 bind to recombinant human BCMA, with similar affinity as
the chimeric molecule. Data generated from this experiment are
presented in table 10.
TABLE-US-00012 TABLE 10 Kinetics analysis of anti-BCMA humanised
molecules against Recombinant Human BCMA Human commercial BCMA Cyno
in house BCMA Sample KD KD name ka kd (nM) ka kd (nM) CA8 1.96E+07
3.50E-04 0.018 6.77E+05 2.99E-04 0.442 humanised J9M1 CA8 4.95E+06
1.74E-04 0.035 7.03E+05 3.24E-04 0.46 humanised J9M0 CA8 3.27E+07
1.18E-03 0.036 1.15E+06 3.49E-04 0.305 Chimera CA8 2.66E+06
1.34E-04 0.05 2.82E+05 3.62E-04 1.284 humanised J8M1 CA8 2.44E+06
1.26E-04 0.052 3.89E+05 4.18E-04 1.076 humanised J8M0 CA8 2.35E+06
1.31E-04 0.056 3.70E+05 3.91E-04 1.057 humanised J7M1 CA8 2.63E+06
1.50E-04 0.057 3.83E+05 5.06E-04 1.324 humanised J8M2 CA8 2.37E+06
1.35E-04 0.057 3.46E+05 4.47E-04 1.293 humanised J7M2 CA8 2.36E+06
1.51E-04 0.064 3.21E+05 3.67E-04 1.143 humanised J7M0 CA8 No Ana-
Bind- 4.88E+05 2.52E-04 0.515 humanised lysable ing J9M2
4.9 BIAcore Analysis of Anti-BCMA CA8 Chimeric and Humanised
Constructs J6M0 and J9M0
[0308] Protein A was immobilised on a CM5 chip (GE Healthcare, Cat
No: BR-1005-30) by primary amine coupling and this surface was then
used to capture the antibody molecules. Recombinant human BCMA (US
Biological, B0410) was used as analyte at 256 nM, 64 nM, 16 nM, 4
nM and 1 nM. Regeneration of the capture surface was carried out
using 50 mM NaOH. All binding curves were double referenced with a
buffer injection (i.e. 0 nM) the data was fitted to the using the
1:1 model inherent to T100 evaluation software. The run was carried
out at 25.degree. C. and 37.degree. C. for experiment 1 and only
37.degree. C. for experiment 2 using HBS-EP as the running
buffer.
[0309] The both runs identified J9M0 as the best molecule in term
of overall affinity to human BCMA. Data generated from this
experiment are presented in table 11.
TABLE-US-00013 TABLE 11 Kinetics analyses of anti-BCMA humanised
molecules against Human BCMA Human commercial BCMA 25.degree. C.
37.degree. C. Experiment 1 Experiment 1 Experiment 2 KD KD KD
Sample ka kd (nM) ka kd (nM) ka kd (nM) J9M0 1.59E+06 3.38E-05
0.021 3.75E+06 1.58E-04 0.042 3.62E+06 1.89E-04 0.052 J6M0 1.01E+06
1.22E-04 0.121 2.12E+06 1.48E-03 0.698 3.78E+06 1.88E-03 0.498
Chimera 1.88E+06 2.63E-04 0.140 1.72E+07 8.72E-04 0.051 1.88E+07
1.04E-03 0.055 CA8
4.10. ProteOn Analysis of New Anti-BCMA Chimeric Constructs
[0310] The initial screen of the new chimeric variants from the
second batch of hybridomas was carried out on the ProteOn XPR36
(Biorad). The method was as follows; Protein A was immobilised on a
GLM chip (Biorad, Cat No: 176-5012) by primary amine coupling,
anti-BCMA variants were then captured on this surface and
recombinant human BCMA (in house material) passed over at 256, 64,
16, 4, 1 nM with a 0 nM injection (i.e. buffer alone) used to
double reference the binding curves, the buffer used is the HBS-EP
buffer. Regeneration of the capture surface was carried out using
50 mM NaOH. The data was fitted to the 1:1 model using the analysis
software inherent to the ProteOn XPR36. The run was carried out at
25.degree. C.
[0311] Data generated from this experiment are presented in table
12.
TABLE-US-00014 TABLE 12 Kinetics analyses of anti-BCMA humanised
molecules against Human BCMA In house human BCMA Sample name ka kd
KD (nM) S332110D07 3.11E+05 3.77E-03 12.100 S332121F02 3.73E+05
6.45E-03 17.300
Example 5
Cell Killing Assays
5.1 ADCC Potencies of Chimeric CA8 and Defucosylated Chimeric CA8
Version in ARH77 Cells Expressing BCMA
[0312] Human natural killer (NK) cells were incubated with europium
labelled ARH77 BCMA transfected target cells (10B5) in the presence
of varying concentrations of antibody at an E:T ratio of 5:1 for 2
hours. Europium release from the target cells was measured and
specific lysis calculated. Result: Chimeric CA8 and defucosylated
chimeric CA8 killed BCMA expressing target cells via ADCC. The
defucosylated chimeric antibody showed more potent ADCC activity,
as measured by a higher percent lysis achieved with all the target
cells tested and a ten-fold lower EC.sub.50 on the high BCMA
expressing target cell line 10B5, compared to the parent chimeric
antibody. See FIGS. 8A and 8B.
5.2 ADCC Activity of CA8 Humanised Antibodies Using ARH77 BCMA
Expressing Target Cells and PBMC as Effectors
[0313] Human PBMC were incubated with europium labelled ARH77 BCMA
transfected target cells (10B5) in the presence of varying
concentrations of humanised versions of CA8 antibody (5 ug/ml to
0.005 ug/ml) at an E:T ratio of 5:1 for 2 hours. Europium release
from the target cells was measured and specific lysis
calculated.
Result:
[0314] Result: All the J5, J6, J7 J8 & J9 series of humanised
variants of CA8 showed ADCC activity against the ARH77 high BCMA
expressing cell line 10B5 in a dose dependent manner. ADCC was at a
similar level as that found in the experiments using chimeric CA8
molecule. See FIG. 9.
5.3 ADCC Potencies of Chimeric S322110F02, S322110D07 and
S307118G03 and Humanised S307118G03 H3L0 Against ARH77 10B5 Cells
Expressing BCMA with Purified NK Cells as Effector Cells
[0315] Human natural killer (NK) target cells were incubated with
europium labelled ARH77 BCMA transfected target cells (10B5) in the
presence of varying concentrations of antibody at an E:T ratio of
5:1 for 2 hours. Europium release from the target cells was
measured and specific lysis calculated. Result: all 4 antibodies
tested showed ADCC activity against ARH77 10B5 cells. See FIG.
10.
5.4 Antibody-Drug Conjugate (ADC) Activity of Chimeric CA8
ADCs.
[0316] Measuring ADC activity of chimeric CA8 antibody, chimeric
CA8-mcMMAF antibody drug conjugates and chimeric CA8-vcMMAE
antibody drug conjugates against human multiple myeloma cell lines.
Multiple Myeloma cell lines were treated with chimeric CA8
antibody-drug conjugates to determine the ADC concentrations
required for growth inhibition and death.
[0317] The antibody drug conjugates tested were added to wells
containing multiple myeloma cells at concentrations ranging from 1
ug/ml to 5 ng/ml. The plates were incubated at 37.degree. C. for 96
hours at which point viable cells were quantitated using Cell titre
Glo. The unconjugated chimeric CA8 antibody showed no significant
growth inhibitory activity at the antibody concentrations that were
tested. The chimeric CA8-mcMMAF antibody-drug conjugate showed
greater growth inhibitory activity than the chimeric CA8-vcMMAE
antibody-drug conjugate in all 4 of the multiple myeloma cell lines
that were tested. See FIG. 11 and Table 13
TABLE-US-00015 TABLE 13 IC.sub.50 values represented in ng/mL for
the chimeric CA8-vcMMAE and the chimeric CA8-mcMMAF antibody-drug
conjugates in 4 different multiple myeloma cell lines IC.sub.50
(ng/mL) Multiple Myeloma CA8 chimera- CA8 chimera- cell lines
vcMMAE mcMMAF NCI-H929 29.5 8.8 U266-B1 18.9 9.7 JJN3 21.8 12.4
OPM2 92.7 58.1
5.5 Measuring Cell Cycle Arrest Activity of Chimeric CA8 Antibody,
Chimeric CA8-mcMMAF Antibody Drug Conjugates and Chimeric
CA8-vcMMAE Antibody Drug Conjugates Against Human Multiple Myeloma
Cell Line H929.
[0318] To determine the mechanism that chimeric CA8 Antibody Drug
Conjugates (ADC's) cause growth inhibition in multiple myeloma
cells, the cell cycle of NCI-H929 cells was monitored by measuring
cellular DNA content through fixed cell propidium iodide staining
at multiple timepoints following chimeric CA8 antibody and chimeric
CA8 ADC treatment.
[0319] At the chimeric CA8 ADC concentration tested (50 ng/mL), the
chimeric CA8-mcMMAF ADC caused significant G2/M cell cycle arrest
(4N DNA content) which peaked at 48 hours. At the later timepoints
48, 72 and 96 hours, treatment with the chimeric CA8-mcMMAF ADC
resulted in accumulation of a cell population with sub-2N DNA
content, which is representative of cell death. At the 50 ng/mL
concentration tested the chimeric CA8-vcMMAE ADC had no significant
effect on G2/M cell cycle arrest or sub-G1 accumulation. See FIG.
12.
5.6 Phospho-Histone-H3 (Thr11) Staining as a Marker for Chimeric
CA8-mcMMAF Antibody Drug Conjugate and Chimeric CA8-vcMMAE Antibody
Drug Conjugate Induced Mitotic Arrest.
[0320] To determine if the accumulation of cells with 4N DNA
content is a specific result of mitotic arrest induced by the
chimeric CA8 ADCs NCI-H929 cells were stained with an
anti-phospho-Histone H3 antibody following treatment with
increasing concentrations of unconjugated chimeric CA8, chimeric
CA8-vcMMAE or chimeric CA8-mcMMAF for 48 hours.
[0321] Treatment with chimeric CA8 ADCs resulted in a
dose-dependent accumulation of NCI-H929 cells that stained positive
for 65eroxidi-Histone H3 (Thr11), a specific marker of mitotic
cells. The chimeric CA8-mcMMAF ADC caused accumulation of
65eroxidi-Histone H3 positive cells at lower concentrations than
the chimeric CA8-vcMMAE ADC. See FIG. 13.
5.7 Measuring Apoptosis in NCI-H929 Cells in Response to Chimeric
CA8 ADCs by Staining for Annexin V.
[0322] To determine if the accumulation of cells with sub-2N DNA
content is a specific result of apoptosis induced by the chimeric
CA8 ADCs, NCI-H929 cells were stained with an anti-Annexin-V
antibody following treatment with increasing concentrations of
unconjugated chimeric CA8, chimeric CA8-vcMMAE or chimeric
CA8-mcMMAF for 48 hours. Treatment with chimeric CA8 ADCs resulted
in a dose-dependent accumulation of NCI-H929 cells that stained
positive for Annexin-V, a specific marker of apoptosis. The
chimeric CA8-mcMMAF ADC caused accumulation of Annexin-V positive
cells at lower concentrations than the chimeric CA8-vcMMAE ADC. See
FIG. 14.
5.8 Antibody-Drug Conjugate (ADC) Activity of Humanised Variants of
CA8 Anti-BCMA Antibody-Drug Conjugates.
[0323] Cells were plated in 96-well plates (4,000 cells per well in
100 uL of RPMI+10% FBS) Naked antibody or ADC was added 6 hours
after cell seeding and plates were incubated for 144 hours. Growth
inhibition in the presence of the antibodies or ADCs was measured
at 144 hours using Cell Titre glo. Data points represent the mean
of triplicate CellTiterGlo measurements. Error bars represent
standard error.
[0324] Multiple Myeloma cell lines NCI-H929 and OPM2 were treated
with humanized CA8 anti-BCMA antibody-drug conjugates to determine
the ADC concentrations required for growth inhibition and death.
The mcMMAF and vcMMAE antibody-drug conjugate forms of these
antibodies showed significant growth inhibitory activity comparable
to that found with the CA8 chimera. Variant J6M0 showed higher
potency than the chimera and data is shown in FIG. 15 in H929 cells
and OPM2 cells. The mcMMAF antibody-drug conjugate showed greater
growth inhibitory activity than the vcMMAE antibody-drug conjugate
for all antibodies in both cell lines tested. Results for all
humanized variants are shown in Table 14.
TABLE-US-00016 TABLE 14 IC.sub.50 values represented in ng/mL for
the anti BCMA antibody-drug conjugates in NCI-H929 and U266-B1
cells NCI-H929 mcMMAF vcMMAE OPM2 vcMMAE Average Average mcMMAF
Average IC50 IC50 Average IC50 IC50 (ng/mL) (ng/mL) (ng/mL) (ng/mL)
CA8 chimera 11.64 37.96 57.04 80.01 CA8 J6M0 5.97 27.67 87.22 121.2
CA8 J6M1 14.6 51.89 205.6 239.9 CA8 J6M2 9.5 39.71 112.9 144.7 CA8
J7M0 18.97 52.25 93.27 127.1 CA8 J7M1 17.87 43.97 95.35 107.5 CA8
J7M2 31.63 55.13 102.6 115.9 CA8 J8M0 15.67 59.94 89.95 132 CA8
J8M1 17.04 46.55 82.96 115.8 CA8 J8M2 15.08 55.98 72.63 124.5 CA8
J9M0 14.95 48.5 58.6 109.8 CA8 J9M1 15.19 55.1 55.88 115 CA8 J9M2
20.87 55.77 80.35 111.7
5.9 Antibody-Drug Conjugate (ADC) Activity of Other Murine
Anti-BCMA Antibody-Drug Conjugates.
[0325] Cells were plated in 96-well plates (4,000 cells per well in
100 uL of RPMI+10% FBS) Antibody or ADC was added 6 hours after
cell seeding and plates were incubated for 144 hours. Growth
inhibition in the presence of the ADCs was measured at 144 hours
using Cell Titre glo. The mean of triplicate CellTiterGlo
measurements are shown. Table 15a and 15b are from experiments
carried out at different times on different series of antibodies.
Multiple Myeloma cell lines NCI-H929 and U266-B1 were used for
antibodies in Table 15a.
[0326] The mcMMAF and vcMMAE antibody-drug conjugate forms of
murine antibodies S322110D07, S332121F02 and S332136E04 showed
significant growth inhibitory activity. The mcMMAF antibody-drug
conjugate showed greater growth inhibitory activity than the vcMMAE
antibody-drug conjugate in all of the murine anti-BCMA antibodies
tested where activity was seen. IC50 figures are shown in Table
15a. See FIG. 16 for dose response curves for these three
antibodies and also S107118G03. Error bars represent standard
error. NCI-H929, U266-B1, JJN3 and OPM2 cells for antibodies in
Table 15b were treated with a different series of murine anti-BCMA
antibody-drug conjugates to determine the ADC concentrations
required for growth inhibition and death. IC50 figures are shown in
Table 15b. All 5 antibodies shown on the table had significant ADC
activity.
TABLE-US-00017 TABLE 15a IC.sub.50 values represented in ng/mL for
the anti BCMA antibody-drug conjugates in NCI-H929 and U266-B1
cells IC50 (ng/mL) NCI-H929 U226-B1 Antibody -vcMMAE -mcMMAF
-vcMMAE -mcMMAF S322110D07 28.4 6.7 53.3 33.3 mIgG1 S332121F02 24.5
7 2.3 2.5 mIgG1 S332126E04 46.8 9.7 27.1 10.6 mIgG1
TABLE-US-00018 TABLE 15b IC.sub.50 values represented in ng/mL for
the anti BCMA antibody-drug conjugates in NCI- H929, U266-B1, JJN3
and OPM2 cells Average IC50 NCI-H929 U266B1 JJN3 OPM2 (ng/mL)
vcMMAE mcMMAF vcMMAE mcMMAF vcMMAE mcMMAF vcMMAE mcMMAF S335115G01
14.9 4.2 38.8 18.5 73.9 45.8 162.4 197.2 S336105A07 17.8 5.1 21.4
9.3 54.2 23.2 95.5 73.7 S335122F05 10.9 4.2 21.1 14.1 29.5 25.5
98.4 128.7 S335106E08 19.2 7.9 36.8 32.6 189.8 214.1 243.9 307.5
S335128A12 86.3 28.3 101.8 104.1 >500 >500 >500
>500
5.10 ADCC Potency of Conjugated, Afucosylated J6M0
(Potelligent)
[0327] Afucosylated J6M0 conjugated to MMAE or MMAF was tested in
ADCC assays using BCMA transfectants to ensure that its ADCC
activity was not compromised by the conjugation. Europium labelled
ARH77-10B5 cells were incubated with various J6M0 WT and
Potelligent BCMA antibodies at concentrations up to 10000 ng/ml for
30 minutes prior to the addition of PBMCs (PBMC: target cell ratio
50:1). Two hours later an aliquot of cell media was sampled and
mixed with enhancement solution. After 30 minutes on a plate
shaker, europium release was monitored on the Victor 2 1420
multi-label reader. Datapoints represent means of triplicate
values. This data is representative of 2 experiments.
[0328] There were no significant differences in ADCC potency
between the unconjugated and ADC forms of J6M0 Potelligent. In the
same experiment a wild type version of J6M0 was included to show
how the potency compares to the afucosylated version. As expected,
defucosylation resulted in a lower EC50 and higher maximal lysis.
No lysis was observed with the Fc disabled form of J6M0. (FIG.
17)
5.11 ADCC Potency of Afucosylated J6M0 on MM Cell Lines
[0329] Human PBMC were incubated with multiple myeloma target cells
at an E:T ratio of 50:1 in presence of varying concentrations of
afucosylated (Potelligent) J6M0 The percentage of target cells
remaining in the effector+target cell mixture after 18 hours was
measured by FACS using a fluorescently labelled anti-CD138 antibody
to detect the target cells and the percent lysis calculated. This
is representative of several experiments.
[0330] J6M0 Potelligent antibody showed ADCC activity against all
five multiple myeloma target cell lines tested. This was important
to test since earlier studies were carried out using transfected
cells. Results are shown in FIG. 18. Full dataset with multiple
donors is shown in Table 16 The potencies were all in a similar
range as those found with the transfectants. The ADCC activity was
not directly related to BCMA surface expression on these cell
lines.
TABLE-US-00019 TABLE 16 EC.sub.50 values generated on 13
independent assays using 11 donors (designated A-K) across the five
multiple myeloma cell lines. EC.sub.50 (ng/mL) Donor H929 RPMI 8226
JJN-3 OPM-2 U266 A 1.43 NA 1.64 NA NA B 0.57 NA NA NA NA C 0.73 NA
1.01 NA NA C 1.81 NA NA NA NA A 2.05 NA NA NA NA D NA 4.09 NA NA NA
E NA NA 14.4 NA NA F 2.18 NA NA NA NA G NA NA 26.3 NA NA H 4.79 NA
111.3 NA NA I NA NA 40.1 NA NA J 2.19 20.4 4.89 NA NA K ND ND 4.52
4.15 9.04
Example 6
Xenograft Data
[0331] 6.1 Murine xenografts of human MM cell lines were tested to
ensure that antibody potency detected in vitro can also be
demonstrated in vivo. The cell line selected for xenograft studies
was NCI-H929 which is sensitive to ADC and ADCC killing in vitro.
Studies were carried out in immunocompromised CB.17 SCID mice which
lack T and B cells but maintain NK cells to allow for ADCC
activity. However it should be noted that although human IgG1 can
engage murine Fc receptors, the Potelligent enhancement does not
improve the affinity as it does with human Fc receptors.
6.2 Impact of Unconjugated and MMAE or MMAF Conjugated J6M0 on
NCI-H929 Tumour Growth.
[0332] In order to independently analyze both the ADCC and ADC
activities of J6M0 we tested J6M0 antibody in the presence and
absence of MMAF or MMAE conjugation. By testing the unconjugated
J6M0, any anti-tumour effects could be attributed to some
combination of ADDC and functional inhibitory activity.
[0333] Mice with NCI-H929 tumours that had reached a volume of 200
mm.sup.3 on average were treated with a human IgG1 control or the
J6M0 antibody (unconjugated, MMAE or MMAF) twice weekly at a dose
of 50 ug or 100 ug, for 2 weeks. Results from this study show that
a 100 ug dose of the J6MO-MMAF conjugate resulted in elimination of
tumours in those mice which have completed the dosing. The
J6MO-MMAF mice were maintained for 40 days after the last dose with
no recurrence of tumour occurring. These results from this
experiment demonstrate that MMAF conjugation had increased
anti-tumour activity over both unconjugated J6M0 antibody and
J6MO-MMAE conjugate See FIG. 19.
Example 7
Evaluation of Soluble BCMA Levels from MM Patient Serum
[0334] 7.1 It is currently unknown whether BCMA is present
extracellularly and can be detected in the blood. In this work, we
determined the serum level of human BCMA from MM patients. Serum
samples from 54 MM and plasma cell dyscrasia patients and 20 normal
control samples were analyzed by ELISA. Human Subject Approval was
obtained from Western Institutional Review Board.
7.2 Assessment of Serum Human BCMA Levels
[0335] Blood, from patients and normal controls in the clinic, were
collected in serum collection tubes. MM patient samples were from a
variety of stages (progressive disease, remission, relapsed, newly
diagnosed, and others). The Blood samples were spun at 10,000 rpm
for 10 minutes and serum transferred into sterile micro-centrifuge
plastic tubes.
[0336] A Human BCMA/TNFRSF17 ELISA kit from R& D Systems
(catalog # DY193E) which measures soluble human BCMA levels was
used to detect BCMA following the standard protocol supplied with
the kit.
[0337] Briefly, 96 well micro-plates were coated with 100 ul per
well capture antibody and incubated overnight at 4.degree. C. The
plates were washed three times with wash buffer (0.05% Tween 20 in
PBS, pH 7.2) and blocked with 300 ul of 1% BSA in PBS at room
temperature for 2 hours. The plates were washed three times with
washing buffer. 100 ul of serum sample or standard was added into
each well and incubated for 2 hours at room temperature. The plates
were washed three times with washing buffer and then 100 ul of the
detection antibody was added to each well and incubated 2 hours at
room temperature. 100 ul of Streptavidin-HRP was added in each well
after washing plates three times and incubated in dark room for 20
minutes. The plates were washed three times and added 50 ul stop
solution and then determined by micro-plate reader with 570 nM
wavelength.
[0338] A series of assays were carried out in order to determine
the serum dilution factor appropriate for the levels of BCMA which
were present. A dilution factor of 1:500 was found to be suitable
for the majority of samples and is the dilution factor used in the
data shown in FIG. 20. The full data set is shown in Table 17.
[0339] Patient and normal control serum samples diluted and run in
triplicates had BCMA levels determined. The serum levels of BCMA
were significantly elevated in the sera from MM patients compared
with normal controls in this study. When the disease subset was
divided further there was a trend towards elevated serum levels of
BCMA in the sera from progressing MM patients compared with those
in remission. This is the first report identifying serum BCMA in
any human disease and suggests that these levels may be a novel
biomarker for monitoring disease status and therapeutic response of
MM patients and for other patients with plasma cell mediated
diseases.
TABLE-US-00020 TABLE 17 Figures represent serum concentration of
soluble BCMA in ng/ml calculated from samples diluted at 1/50,
1/500 and 1/5000. P values were calculated using the one tailed
T-Test and 95% significance values are below the table. Myeloma:
Myeloma: Myeloma: Myeloma: Other Plasma Cell Normal Progressive
Stable Remission Other MGUS Dyscrasias 1-5000 Mean 14.130 500.804
154.762 151.201 94.457 84.912 22.838 1-500 Triplicate Mean 15.901
215.877 81.135 43.294 97.584 53.894 22.838 1-500 Single Mean 16.620
207.028 61.576 42.796 71.372 40.623 14.099 1-50 Trial 1 Mean 25.568
129.544 41.983 40.507 65.120 42.067 51.650 1-50 Trial 2 Mean 17.160
119.220 34.567 34.264 54.780 26.333 51.650 P-Values (One Tailed
T-Test, 95% Significance) ~1-500 Single Normal vs Progressive: p =
.0010* Progressive vs Remission: p = .0146* ~1-500 Triplicate
Normal vs Progressive: p = .0004* Progressive vs Remission: p =
.0091* ~1-50 Trial 1 Normal vs Progressive: p = .0171* Progressive
vs Remission: p = .0777 ~1-50 Trial 2 Normal vs Progressive: p =
.0184* Progressive vs Remission: p = .0876 *shows significance
TABLE-US-00021 TABLE C Sequence Summary Polynucleotide Description
Amino acid sequence sequence CA8 CDRH1 SEQ. I.D. NO: 1 n/a CA8
CDRH2 SEQ. I.D. NO: 2 n/a CA8 CDRH3 SEQ. I.D. NO: 3 n/a CA8 CDRL1
SEQ. I.D. NO: 4 n/a CA8 CDRL2 SEQ. I.D. NO: 5 n/a CA8 CDRL3 SEQ.
I.D. NO: 6 n/a CA8 V.sub.H domain (murine) SEQ. I.D. NO: 7 SEQ.
I.D. NO: 8 CA8 V.sub.L domain (murine) SEQ. I.D. NO: 9 SEQ. I.D.
NO: 10 CA8 Humanised V.sub.H J0 SEQ. I.D. NO: 11 SEQ. I.D. NO: 12
CA8 Humanised V.sub.H J1 SEQ. I.D. NO: 13 SEQ. I.D. NO: 14 CA8
Humanised V.sub.H J2 SEQ. I.D. NO: 15 SEQ. I.D. NO: 16 CA8
Humanised V.sub.H J3 SEQ. I.D. NO: 17 SEQ. I.D. NO: 18 CA8
Humanised V.sub.H J4 SEQ. I.D. NO: 19 SEQ. I.D. NO: 20 CA8
Humanised V.sub.H J5 SEQ. I.D. NO: 21 SEQ. I.D. NO: 22 CA8
Humanised V.sub.H J6 SEQ. I.D. NO: 23 SEQ. I.D. NO: 24 CA8
Humanised V.sub.H J7 SEQ. I.D. NO: 25 SEQ. I.D. NO: 26 CA8
Humanised V.sub.H J8 SEQ. I.D. NO: 27 SEQ. I.D. NO: 28 CA8
Humanised V.sub.H J9 SEQ. I.D. NO: 29 SEQ. I.D. NO: 30 CA8
Humanised V.sub.L M0 SEQ. I.D. NO: 31 SEQ. I.D. NO: 32 CA8
Humanised V.sub.L M1 SEQ. I.D. NO: 33 SEQ. I.D. NO: 34 CA8
Humanised V.sub.L M2 SEQ. I.D. NO: 35 SEQ. I.D. NO: 36 Human BCMA
SEQ. I.D. NO: 37 SEQ. I.D. NO: 38 CD33-hBCMA ECD (1-53) TEV-Fc
Human BCMA SEQ. I.D. NO: 39 SEQ. I.D. NO: 40 CD33-hBCMA ECD (4-53)
TEV-Fc Cyno BCMA SEQ. I.D. NO: 41 SEQ. I.D. NO: 42 CD33 cyno BCMA
ECD (4-52) TEV-Fc CA8 J0 Humanised heavy chain SEQ. I.D. NO: 43
SEQ. I.D. NO: 44 CA8 J1 Humanised heavy chain SEQ. I.D. NO: 45 SEQ.
I.D. NO: 46 CA8 J2 Humanised heavy chain SEQ. I.D. NO: 47 SEQ. I.D.
NO: 48 CA8 J3 Humanised heavy chain SEQ. I.D. NO: 49 SEQ. I.D. NO:
50 CA8 J4 Humanised heavy chain SEQ. I.D. NO: 51 SEQ. I.D. NO: 52
CA8 J5 Humanised heavy chain SEQ. I.D. NO: 53 SEQ. I.D. NO: 54 CA8
J6 Humanised heavy chain SEQ. I.D. NO: 55 SEQ. I.D. NO: 56 CA8 J7
Humanised heavy chain SEQ. I.D. NO: 57 SEQ. I.D. NO: 58 CA8 J8
Humanised heavy chain SEQ. I.D. NO: 59 SEQ. I.D. NO: 60 CA8 J9
Humanised heavy chain SEQ. I.D. NO: 61 SEQ. I.D. NO: 62 CA8 M0
Humanised light chain SEQ. I.D. NO: 63 SEQ. I.D. NO: 64 CA8 M1
Humanised light chain SEQ. I.D. NO: 65 SEQ. I.D. NO: 66 CA8 M2
Humanised light chain SEQ. I.D. NO: 67 SEQ. I.D. NO: 68 S307118G03
V.sub.H domain (murine) SEQ. I.D. NO: 69 SEQ. I.D. NO: 70
S307118G03 V.sub.L domain (murine) SEQ. I.D. NO: 71 SEQ. I.D. NO:
72 S307118G03 heavy chain (chimeric) SEQ. I.D. NO: 73 SEQ. I.D. NO:
74 S307118G03 light chain(chimeric) SEQ. I.D. NO: 75 SEQ. I.D. NO:
76 S307118G03 Humanised V.sub.H H0 SEQ. I.D. NO: 77 SEQ. I.D. NO:
78 S307118G03 Humanised V.sub.H H1 SEQ. I.D. NO: 79 SEQ. I.D. NO:
80 S307118G03 humanised V.sub.H H2 SEQ. I.D. NO: 81 SEQ. I.D. NO:
82 S307118G03 humanised V.sub.H H3 SEQ. I.D. NO: 83 SEQ. I.D. NO:
84 S307118G03 humanised V.sub.H H4 SEQ. I.D. NO: 85 SEQ. I.D. NO:
86 S307118G03 humanised V.sub.H H5 SEQ. I.D. NO: 87 SEQ. I.D. NO:
88 S307118G03 humanised V.sub.L L0 SEQ. I.D. NO: 89 SEQ. I.D. NO:
90 S307118G03 humanised V.sub.L L1 SEQ. I.D. NO: 91 SEQ. I.D. NO:
92 S307118G03 CDRH1 SEQ. I.D. NO: 93 S307118G03 CDRH2 SEQ. I.D. NO:
94 S307118G03 CDRH3 SEQ. I.D. NO: 95 S307118G03 CDRL1 SEQ. I.D. NO:
96 S307118G03 CDRL2 SEQ. I.D. NO: 97 S307118G03 CDRL3 SEQ. I.D. NO:
98 S307118G03 humanised H5 CDRH3 SEQ. I.D. NO: 99 S307118G03 H0
Humanised heavy SEQ. I.D. NO: 100 SEQ. I.D. NO: 101 chain
S307118G03 H1 humanised heavy SEQ. I.D. NO: 102 SEQ. I.D. NO: 103
chain S307118G03 H2 humanised heavy SEQ. I.D. NO: 104 SEQ. I.D. NO:
105 chain S307118G03 H3 humanised heavy SEQ. I.D. NO: 106 SEQ. I.D.
NO: 107 chain S307118G03 H4 humanised heavy SEQ. I.D. NO: 108 SEQ.
I.D. NO: 109 chain S307118G03 H5 humanised heavy SEQ. I.D. NO: 110
SEQ. I.D. NO: 111 chain S307118G03 L0 humanised light chain SEQ.
I.D. NO: 112 SEQ. I.D. NO: 113 S307118G03 L1 humanised light chain
SEQ. I.D. NO: 114 SEQ. I.D. NO: 115 S332121F02 murine variable
heavy SEQ. I.D. NO: 116 SEQ. I.D. NO: 117 chain S332121F02 chimeric
variable heavy SEQ. I.D. NO: 118 SEQ. I.D. NO: 119 chain S332121F02
murine variable light chain SEQ. I.D. NO: 120 SEQ. I.D. NO: 121
S332121F02 chimeric variable light SEQ. I.D. NO: 122 SEQ. I.D. NO:
123 chain S322110D07 murine variable heavy SEQ. I.D. NO: 124 SEQ.
I.D. NO: 125 chain S322110D07 chimeric heavy chain SEQ. I.D. NO:
126 SEQ. I.D. NO: 127 S322110D07 murine variable light SEQ. I.D.
NO: 128 SEQ. I.D. NO: 129 chain S322110D07 chimeric light chain
SEQ. I.D. NO: 130 SEQ. I.D. NO: 131 S332126E04 murine variable
heavy SEQ. I.D. NO: 132 SEQ. I.D. NO: 133 chain S332126E04 Chimeric
heavy chain SEQ. I.D. NO: 134 SEQ. I.D. NO: 135 S332126E04 murine
variable light chain SEQ. I.D. NO: 136 SEQ. I.D. NO: 137 S332126E04
Chimeric light chain SEQ. I.D. NO: 138 SEQ. I.D. NO: 139 S336105A07
murine variable heavy SEQ. I.D. NO: 140 SEQ. I.D. NO: 141 chain
S336105A07 Chimeric heavy chain SEQ. I.D. NO: 142 SEQ. I.D. NO: 143
S336105A07 murine variable light chain SEQ. I.D. NO: 144 SEQ. I.D.
NO: 145 S336105A07 chimeric light chain SEQ. I.D. NO: 146 SEQ. I.D.
NO: 147 S335115G01 murine variable heavy SEQ. I.D. NO: 148 SEQ.
I.D. NO: 149 chain S335115G01 Chimeric heavy chain SEQ. I.D. NO:
150 SEQ. I.D. NO: 151 S335115G01 murine variable light chain SEQ.
I.D. NO: 152 SEQ. I.D. NO: 153 S335115G01 Chimeric light chain SEQ.
I.D. NO: 154 SEQ. I.D. NO: 155 S335122F05 murine variable heavy
SEQ. I.D. NO: 156 SEQ. I.D. NO: 158 chain S335122F05 Chimeric heavy
chain SEQ. I.D. NO: 158 SEQ. I.D. NO: 159 S335122F05 murine
variable light chain SEQ. I.D. NO: 160 SEQ. I.D. NO: 161 S335122F05
Chimeric light chain SEQ. I.D. NO: 162 SEQ. I.D. NO: 163 S332121F02
CDRH1 SEQ. I.D. NO: 164 S332121F02 CDRH2 SEQ. I.D. NO: 165
S332121F02 CDRH3 SEQ. I.D. NO: 166 S332121F02 CDRL1 SEQ. I.D. NO:
167 S332121F02 CDRL2 SEQ. I.D. NO: 168 S332121F02 CDRL3 SEQ. I.D.
NO: 169 S322110D07 CDRH1 SEQ. I.D. NO: 170 S322110D07 CDRH2 SEQ.
I.D. NO: 171 S322110D07 CDRH3 SEQ. I.D. NO: 172 S322110D07CDRL1
SEQ. I.D. NO: 173 S322110D07 CDRL2 SEQ. I.D. NO: 174 S322110D07
CDRL3 SEQ. I.D. NO: 175 S332126E04CDRH1 SEQ. I.D. NO: 176
S332126E04 CDRH2 SEQ. I.D. NO: 177 S332126E04 CDRH3 SEQ. I.D. NO:
178 S332126E04 CDRL1 SEQ. I.D. NO: 179 S332126E04 CDRL2 SEQ. I.D.
NO: 180 S332126E04 CDRL3 SEQ. I.D. NO: 181 S336105A07 CDRH1 SEQ.
I.D. NO: 182 S336105A07 CDRH2 SEQ. I.D. NO: 183 S336105A07 CDRH3
SEQ. I.D. NO: 184 S336105A07 CDRL1 SEQ. I.D. NO: 185 S336105A07
CDRL2 SEQ. I.D. NO: 186 S336105A07 CDRL3 SEQ. I.D. NO: 187
S335115G01 CDRH1 SEQ. I.D. NO: 188 S335115G01 CDRH2 SEQ. I.D. NO:
189 S335115G01 CDRH3 SEQ. I.D. NO: 190 S335115G01 CDRL1 SEQ. I.D.
NO: 191 S335115G01 CDRL2 SEQ. I.D. NO: 192 S335115G01 CDRL3 SEQ.
I.D. NO: 193 S335122F05 CDRH1 SEQ. I.D. NO: 194 S335122F05 CDRH2
SEQ. I.D. NO: 195 S335122F05 CDRH3 SEQ. I.D. NO: 196 S335122F05
CDRL1 SEQ. I.D. NO: 197 S335122F05 CDRL2 SEQ. I.D. NO: 198
S335122F05 CDRL3 SEQ. I.D. NO: 199
TABLE-US-00022 SEQUENCE LISTING CA8 CDRH1 SEQ ID 1 NYWMH CA8 CDRH2
SEQ ID 2 ATYRGHSDTYYNQKFKG CA8 CDRH3 SEQ ID 3 GAIYNGYDVLDN CA8
CDRL1 SEQ ID 4 SASQDISNYLN CA8 CDRL2 SEQ ID 5 YTSNLHS CA8 CDRL3 SEQ
ID 6 QQYRKLPWT CA8 V.sub.H domain (murine) SEQ ID 7
EVQLQQSGAVLARPGASVKMSCKGSGYTFTNYWMHVVVKQRPGQGLEWIGATYRGHSDTYYNQKF
KGKAKLTAVTSTSTAYMELSSLTNEDSAVYYCTRGAIYNGYDVLDNWGQGTLVTVSS CA8
V.sub.H domain (murine) (Polynucleotide) SEQ ID 8
GAGGTGCAGCTGCAGCAGAGCGGCGCCGTGCTGGCCAGGCCCGGAGCTAGCGTGAAGATGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAAACAGAGGCCCGG
CCAGGGACTGGAGTGGATCGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCAAGGCCAAGCTGACCGCCGTGACCTCAACCAGCACCGCCTACATGGAACTGAG
CAGCCTGACCAACGAGGACAGCGCCGTCTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAATTGGGGCCAGGGAACACTAGTGACCGTGTCCAGC CA8 V.sub.L
domain (murine) SEQ ID 9
DIQLTQTTSSLSASLGDRVTISCSASQDISNYLNWYQQKPDGTVELVIYYTSNLHSGVPSRFSGSGSG
TDYSLTIGYLEPEDVATYYCQQYRKLPWTFGGGSKLEIKR CA8 V.sub.L domain
(murine) (Polynucleotide) SEQ ID 10
GATATCCAGCTGACCCAGACCACAAGCAGCCTGAGCGCCTCCCTGGGCGACAGGGTGACCATT
AGCTGCAGCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGACGGC
ACCGTGGAGCTCGTGATCTACTACACCTCCAACCTGCACAGCGGCGTGCCCAGCAGGTTCTCTG
GCAGCGGCAGCGGCACCGACTACAGCCTGACCATCGGCTATCTGGAGCCCGAGGACGTCGCCA
CCTACTACTGCCAGCAGTACAGGAAGCTGCCCTGGACCTTCGGCGGAGGCTCTAAGCTGGAGA
TTAAGCGT CA8 Humanised V.sub.H J0 SEQ ID 11
QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYNGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J0 (Polynucleotide) SEQ ID 12
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCGGCACCTTCAGCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCG
GACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGA
AGTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGA
GCAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACAACGGCT
ACGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J1 SEQ ID 13
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYNGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J1 (Polynucleotide) SEQ ID 14
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J2 SEQ ID 15
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCTRGAIYNGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J2 (Polynucleotide) SEQ ID 16
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J3 SEQ ID 17
QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADTSTSTAYMELSSLRSEDTAVYYCTRGAIYNGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J3 (Polynucleotide) SEQ ID 18
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J4 SEQ ID 19
QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWIGATYRGHSDTYYNQKF
KGRATLTADTSTSTAYMELSSLRSEDTAVYYCTRGAIYNGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J4 (Polynucleotide) SEQ ID 20
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATCGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGCGACCCTCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J5 SEQ ID 21
QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADTSTSTAYMELSSLRSEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J5 (Polynucleotide) SEQ ID 22
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J6 SEQ ID 23
QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J6 (Polynucleotide) SEQ ID 24
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCGGCACCTTCAGCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCG
GACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGA
AGTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGA
GCAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACGACGGCT
ACGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J7 SEQ ID 25
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J7 (Polynucleotide) SEQ ID 26
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J8 SEQ ID 27
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J8 (Polynucleotide) SEQ ID 28
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.H J9 SEQ ID 29
QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWIGATYRGHSDTYYNQKF
KGRATLTADTSTSTAYMELSSLRSEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSS CA8
Humanised V.sub.H J9 (Polynucleotide) SEQ ID 30
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATCGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGCGACCCTCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC CA8 Humanised
V.sub.L M0 SEQ ID 31
DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKR CA8 Humanised V.sub.L M0
(Polynucleotide) SEQ ID 32
GACATCCAGATGACCCAGAGCCCTAGCTCACTGAGCGCCAGCGTGGGCGACAGGGTGACCATT
ACCTGCTCCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCCAAGCTGCTGATCTACTACACCTCCAACCTGCACTCCGGCGTGCCCAGCAGGTTCAGCG
GAAGCGGCAGCGGCACCGATTTCACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGCCA
CCTACTACTGCCAGCAGTACAGGAAGCTCCCCTGGACTTTCGGCCAGGGCACCAAACTGGAGAT
CAAGCGT CA8 Humanised V.sub.L M1 SEQ ID 33
DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGSGS
GTDYTLTISSLQPEDFATYYCQQYRKLPVVTFGQGTKLEIKR CA8 Humanised V.sub.L M1
(Polynucleotide) SEQ ID 34
GACATCCAGATGACCCAGAGCCCTAGCTCACTGAGCGCCAGCGTGGGCGACAGGGTGACCATT
ACCTGCTCCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCCAAGCTGCTGATCTACTACACCTCCAACCTGCACTCCGGCGTGCCCAGCAGGTTCAGCG
GAAGCGGCAGCGGCACCGATTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGCCA
CCTACTACTGCCAGCAGTACAGGAAGCTCCCCTGGACTTTCGGCCAGGGCACCAAACTGGAGAT
CAAGCGT CA8 Humanised V.sub.L M2 SEQ ID 35
DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPELVIYYTSNLHSGVPSRFSGSGSG
TDYTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKR CA8 Humanised V.sub.L M2
(Polynucleotide) SEQ ID 36
GACATCCAGCTGACCCAGAGCCCTAGCTCACTGAGCGCCAGCGTGGGCGACAGGGTGACCATT
ACCTGCTCCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCCGAGCTGGTGATCTACTACACCTCCAACCTGCACTCCGGCGTGCCCAGCAGGTTCAGC
GGAAGCGGCAGCGGCACCGATTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGCC
ACCTACTACTGCCAGCAGTACAGGAAGCTCCCCTGGACTTTCGGCCAGGGCACCAAACTGGAGA
TCAAGCGT Human BCMA CD33-hBCMA ECD (1-53) TEV-Fc SEQ ID 37
MPLLLLLPLLWAGALAMLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKG
TNSGENLYFQGDPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDP
EVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK
AKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF
LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Human BCMA CD33-hBCMA
ECD (1-53) TEV-Fc (Polynucleotide) SEQ ID 38
ATGCCGCTGCTGCTACTGCTGCCCCTGCTGTGGGCAGGGGCGCTAGCTATGCTGCAGATGGCC
GGCCAGTGCAGCCAGAACGAGTACTTCGACAGCCTGCTGCACGCCTGCATCCCCTGCCAGCTG
AGATGCAGCAGCAACACACCTCCTCTGACCTGCCAGAGATACTGCAACGCCAGCGTGACCAACA
GCGTGAAGGGCACCAACTCCGGAGAGAACCTGTACTTCCAAGGGGATCCCAAATCTTGTGACAA
AACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTC
CCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTG
GACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCAT
AATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTC
ACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC
TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGT
ACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCA
AAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACT
ACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGT
GGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCA
CAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA Human BCMA CD33-hBCMA
ECD (4-53) TEV-Fc SEQ ID 39
MPLLLLLPLLWAGALAMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNS
GENLYFQGDPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS
KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Human BCMA CD33-hBCMA ECD
(4-53) TEV-Fc (Polynucleotide) SEQ ID 40
ATGCCGCTGCTGCTACTGCTGCCCCTGCTGTGGGCAGGGGCGCTAGCTATGGCCGGCCAGTGC
AGCCAGAACGAGTACTTCGACAGCCTGCTGCACGCCTGCATCCCCTGCCAGCTGAGATGCAGC
AGCAACACACCTCCTCTGACCTGCCAGAGATACTGCAACGCCAGCGTGACCAACAGCGTGAAGG
GCACCAACTCCGGAGAGAACCTGTACTTCCAAGGGGATCCCAAATCTTGTGACAAAACTCACAC
ATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAA
CCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC
CACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAG
ACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTG
CACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCC
CCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTG
CCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCT
ATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCA
CGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAG
CAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTAC
ACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA Cynomolgous BCMA CD33 cyno BCMA
ECD (4-52) TEV-Fc SEQ ID 41
MPLLLLLPLLWAGALAMARQCSQNEYFDSLLHDCKPCQLRCSSTPPLTCQRYCNASMTNSVKGMNS
GENLYFQGDPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG
QPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYS
KLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Cynomolgous BCMA CD33 cyno
BCMA ECD (4-52) TEV-Fc (Polynucleotide) SEQ ID 42
ATGCCGCTGCTGCTACTGCTGCCCCTGCTGTGGGCAGGGGCGCTAGCTATGGCCAGACAGTGC
AGCCAGAACGAGTACTTCGACAGCCTGCTGCACGACTGCAAGCCCTGCCAGCTGAGATGCAGC
AGCACACCTCCTCTGACCTGCCAGAGATACTGCAACGCCAGCATGACCAACAGCGTGAAGGGCA
TGAACTCCGGAGAGAACCTGTACTTCCAAGGGGATCCCAAATCTTGTGACAAAACTCACACATGC
CCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCA
AGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACG
AAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAA
GCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCA
GGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATC
GAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAGCCACAGGTGTACACCCTGCCCCCA
TCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCA
GCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTC
CCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTG
GCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAG
AAGAGCCTCTCCCTGTCTCCGGGTAAA CA8 JO Humanised heavy chain SEQ ID 43
QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYNGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J0
Humanised heavy chain (Polynucleotide) SEQ ID 44
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCGGCACCTTCAGCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCG
GACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGA
AGTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGA
GCAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACAACGGCT
ACGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J1 Humanised heavy chain SEQ ID
45 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYNGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J1
Humanised heavy chain (Polynucleotide) SEQ ID 46
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J2 Humanised heavy chain SEQ ID
47 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCTRGAIYNGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J2
Humanised heavy chain (Polynucleotide) SEQ ID 48
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J3 Humanised heavy chain SEQ ID
49 QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADTSTSTAYMELSSLRSEDTAVYYCTRGAIYNGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J3
Humanised heavy chain (Polynucleotide) SEQ ID 50
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J4 Humanised heavy chain SEQ ID
51
QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWIGATYRGHSDTYYNQKF
KGRATLTADTSTSTAYMELSSLRSEDTAVYYCTRGAIYNGYDVLDNWGQGTLVTVSSASTKGPSVFP
LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG
TQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP
PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J4
Humanised heavy chain (Polynucleotide) SEQ ID 52
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATCGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGCGACCCTCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACAACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J5 Humanised heavy chain SEQ ID
53 QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADTSTSTAYMELSSLRSEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK
CA8 J5 Humanised heavy chain (Polynucleotide) SEQ ID 54
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J6 Humanised heavy chain SEQ ID
55 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J6
Humanised heavy chain (Polynucleotide) SEQ ID 56
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCGGCACCTTCAGCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCG
GACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGA
AGTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGA
GCAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACGACGGCT
ACGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J7 Humanised heavy chain SEQ ID
57 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHVVVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J7
Humanised heavy chain (Polynucleotide) SEQ ID 58
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCGCCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J8 Humanised heavy chain SEQ ID
59 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQK
FKGRVTITADKSTSTAYMELSSLRSEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVF
PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL
GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC
VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTT
PPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK CA8 J8
Humanised heavy chain (Polynucleotide) SEQ ID 60
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATGGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 J9 Humanised heavy chain SEQ ID
61
QVQLVQSGAEVKKPGSSVKVSCKGSGYTFTNYWMHVVVRQAPGQGLEWIGATYRGHSDTYYNQKF
KGRATLTADTSTSTAYMELSSLRSEDTAVYYCTRGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVFP
LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG
TQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV
VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP
PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGK CA8 J9
Humanised heavy chain (Polynucleotide) SEQ ID 62
CAGGTGCAGCTGGTCCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCAGCTCCGTGAAAGTGAG
CTGCAAGGGCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAGGCAGGCCCCCGG
ACAGGGCCTGGAGTGGATCGGCGCCACCTACAGGGGCCACAGCGACACCTACTACAACCAGAA
GTTCAAGGGCCGGGCGACCCTCACCGCCGACACGAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTCAGGAGCGAGGACACCGCTGTGTATTACTGCACCAGGGGCGCCATCTACGACGGCTA
CGACGTGCTGGACAACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGG
GCCCCAGCGTGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTG
GGCTGCCTGGTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTG
ACCAGCGGCGTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGC
GTGGTGACCGTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAG
CCCAGCAACACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGC
CCCCCCTGCCCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCT
AAGGACACCCTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCAC
GAGGACCCTGAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACC
AAGCCCAGGGAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCAC
CAGGATTGGCTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTA
TCGAGAAAACCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCC
CTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCC
CAGCGACATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCC
CCCTGTGCTGGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGA
TGGCAGCAGGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACC
CAGAAGAGCCTGAGCCTGTCCCCTGGCAAG CA8 MO Humanised light chain SEQ ID
63
DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL
NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC CA8 MO Humanised light chain (Polynucleotide) SEQ ID
64 GACATCCAGATGACCCAGAGCCCTAGCTCACTGAGCGCCAGCGTGGGCGACAGGGTGACCATT
ACCTGCTCCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCCAAGCTGCTGATCTACTACACCTCCAACCTGCACTCCGGCGTGCCCAGCAGGTTCAGCG
GAAGCGGCAGCGGCACCGATTTCACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGCCA
CCTACTACTGCCAGCAGTACAGGAAGCTCCCCTGGACTTTCGGCCAGGGCACCAAACTGGAGAT
CAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAG
CGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTG
GAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCA
AGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACA
AGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACC
GGGGCGAGTGC CA8 M1 Humanised light chain SEQ ID 65
DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGSGS
GTDYTLTISSLQPEDFATYYCQQYRKLPVVTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL
NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC CA8 M1 Humanised light chain (Polynucleotide) SEQ ID
66 GACATCCAGATGACCCAGAGCCCTAGCTCACTGAGCGCCAGCGTGGGCGACAGGGTGACCATT
ACCTGCTCCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCCAAGCTGCTGATCTACTACACCTCCAACCTGCACTCCGGCGTGCCCAGCAGGTTCAGCG
GAAGCGGCAGCGGCACCGATTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGCCA
CCTACTACTGCCAGCAGTACAGGAAGCTCCCCTGGACTTTCGGCCAGGGCACCAAACTGGAGAT
CAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAG
CGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTG
GAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCA
AGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACA
AGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACC
GGGGCGAGTGC CA8 M2 Humanised light chain SEQ ID 67
DIQLTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPELVIYYTSNLHSGVPSRFSGSGSG
TDYTLTISSLQPEDFATYYCQQYRKLPVVTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN
NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS
PVTKSFNRGEC CA8 M2 Humanised light chain (Polynucleotide) SEQ ID 68
GACATCCAGCTGACCCAGAGCCCTAGCTCACTGAGCGCCAGCGTGGGCGACAGGGTGACCATT
ACCTGCTCCGCCAGCCAGGACATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCCGAGCTGGTGATCTACTACACCTCCAACCTGCACTCCGGCGTGCCCAGCAGGTTCAGC
GGAAGCGGCAGCGGCACCGATTACACCCTGACCATCTCCAGCCTGCAGCCCGAGGACTTCGCC
ACCTACTACTGCCAGCAGTACAGGAAGCTCCCCTGGACTTTCGGCCAGGGCACCAAACTGGAGA
TCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGA
GCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGT
GGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGC
AAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCAC
AAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAAC
CGGGGCGAGTGC S307118G03 mouse variable heavy SEQ ID 69
EVQLQQSGPELVKPGASVKISCKASGYTFTDYYMKWVKQSHGKSLEWIGEIYPNNGGITYNQKFKGK
ATLTVDKSSSTAYMELRSLTSEDSAVYYCANGYEFVYWGQGTLVTVSA S307118G03 mouse
variable heavy (DNA sequence) SEQ ID 70
GAGGTCCAGTTGCAACAATCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCT
GTAAGGCTTCTGGATACACATTCACTGACTACTACATGAAGTGGGTGAAGCAGAGCCATGGAAA
GAGCCTTGAGTGGATTGGAGAGATTTATCCTAATAATGGTGGTATTACCTACAACCAGAAGTTCA
AGGGCAAGGCCACATTGACTGTAGACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCT
GACATCTGAGGACTCTGCAGTCTATTACTGTGCAAATGGTTACGAGTTTGTTTACTGGGGCCAAG
GGACTCTGGTCACTGTCTCTGCA S307118G03 mouse variable light SEQ ID 71
DIQMTQTASSLSASLGDRVTISCSASQGISNYLNWYQQKPDGTVKLLIYYTSSLHSGVPSRFSGSGSG
TDYSLTISNLEPEDIATYYCQQYSKLPVVTFGGGTKLEIKR S307118G03 mouse variable
light (DNA sequence) SEQ ID 72
GATATCCAGATGACACAGACTGCATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCA
GTTGCAGTGCAAGTCAGGGCATTAGCAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACT
GTTAAACTCCTGATCTATTACACATCAAGTTTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAG
TGGGTCTGGGACAGATTATTCTCTCACCATCAGCAACCTGGAACCTGAAGATATTGCCACTTACT
ATTGTCAGCAGTATAGTAAGCTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACG G
S307118G03 chimeric heavy chain SEQ ID 73
EVQLQQSGPELVKPGASVKISCKASGYTFTDYYMKWVKQSHGKSLEWIGEIYPNNGGITYNQKFKGK
ATLTVDKSSSTAYMELRSLTSEDSAVYYCANGYEFVYWGQGTLVTVSAAKTTAPSVFPLAPSSKSTS
GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN
HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED
PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS
KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF
FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 chimeric
heavy chain (DNA sequence) SEQ ID 74
GAGGTCCAGTTGCAACAATCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCT
GTAAGGCTTCTGGATACACATTCACTGACTACTACATGAAGTGGGTGAAGCAGAGCCATGGAAA
GAGCCTTGAGTGGATTGGAGAGATTTATCCTAATAATGGTGGTATTACCTACAACCAGAAGTTCA
AGGGCAAGGCCACATTGACTGTAGACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCT
GACATCTGAGGACTCTGCAGTCTATTACTGTGCAAATGGTTACGAGTTTGTTTACTGGGGCCAAG
GGACTCTGGTCACTGTCTCTGCAGCCAAAACAACAGCCCCCAGCGTGTTCCCCCTGGCCCCCAG
CAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACTACTTCCCCGA
ACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCTTCCCCGCCGT
GCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCAGCAGCCTGG
GCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGGACAAGAAGGT
GGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCGAGCTGCTGGG
AGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCAGCAGAACCCCC
GAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGTTCAACTGGTAC
GTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGTACAACAGCACC
TACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAGGAGTACAAG
TGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAAGGCCAAGGGCC
AGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGG
TGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGAGCA
ACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGATGGCAGCTTCT
TCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGTGTTCAGCTGCT
CCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTGTCCCCTGGCA AG
S307118G03 chimeric light chain SEQ ID 75
DIQMTQTASSLSASLGDRVTISCSASQGISNYLNWYQQKPDGTVKLLIYYTSSLHSGVPSRFSGSGSG
TDYSLTISNLEPEDIATYYCQQYSKLPVVTFGGGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN
NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS
PVTKSFNRGEC S307118G03 chimeric light chain (DNA sequence) SEQ ID
76 GATATCCAGATGACACAGACTGCATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCA
GTTGCAGTGCAAGTCAGGGCATTAGCAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACT
GTTAAACTCCTGATCTATTACACATCAAGTTTACACTCAGGAGTCCCATCAAGGTTCAGTGGCAG
TGGGTCTGGGACAGATTATTCTCTCACCATCAGCAACCTGGAACCTGAAGATATTGCCACTTACT
ATTGTCAGCAGTATAGTAAGCTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAGCTGAAACG
TACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCAC
CGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGT
GGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACT
CCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGT
ACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCG
AGTGC S307118G03 humanised HO variable heavy SEQ ID 77
QVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYYMKWVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCARGYEFVYWGQGTLVTVSS S307118G03
humanised HO variable heavy (DNA sequence) SEQ ID 78
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCGGCACCTTCAGCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAGGGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGC S307118G03 humanised H1 variable
heavy SEQ ID 79
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCARGYEFVYWGQGTLVTVSS S307118G03
humanised H1 variable heavy (DNA sequence) SEQ ID 80
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAGGGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGC S307118G03 humanised H2 variable
heavy SEQ ID 81
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWMGEIYPNNGG ITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCANGYEFVYWGQGTLVTVSS S307118G03
humanised H2 variable heavy (DNA sequence) SEQ ID 82
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAACGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGC S307118G03 humanised H3 variable
heavy SEQ ID 83
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWIGEIYPNNGGITYNQKFKG
RATLTVDKSTSTAYMELSSLRSEDTAVYYCANGYEFVYWGQGTLVTVSS S307118G03
humanised H3 variable heavy (DNA sequence) SEQ ID 84
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATAGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGCGACCCTCACCGTCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAACGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGC S307118G03 humanised H4 variable
heavy SEQ ID 85
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCADGYEFVYWGQGTLVTVSS S307118G03
humanised H4 variable heavy (DNA sequence) SEQ ID 86
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCGACGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGC S307118G03 humanised H5 variable
heavy SEQ ID 87
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWIGEIYPNNGGITYNQKFKG
RATLTVDKSTSTAYMELSSLRSEDTAVYYCANGYEFDYWGQGTLVTVSS S307118G03
humanised H5 variable heavy (DNA sequence) SEQ ID 88
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATAGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGCGACCCTCACCGTCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAACGGCTACGAGTTCGACTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGC S307118G03 humanised L0 variable
light SEQ ID 89
DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQYSKLPVVTFGQGTKLEIKR S307118G03 humanised L0
variable light (DNA sequence) SEQ ID 90
GACATCCAGATGACCCAGAGCCCCTCAAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACTATC
ACCTGCAGCGCCTCCCAGGGCATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCTAAGCTGCTGATCTACTACACCAGCAGCCTGCACAGCGGCGTGCCCAGCAGGTTCTCC
GGCAGCGGCAGCGGAACCGACTTCACCCTGACCATTAGCAGCCTCCAGCCCGAGGACTTCGCC
ACCTACTACTGCCAGCAGTACAGCAAGCTGCCCTGGACCTTCGGCCAGGGCACCAAACTGGAG
ATCAAGCGT S307118G03 humanised L1 variable light SEQ ID 91
DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGS
GTDYTLTISSLQPEDFATYYCQQYSKLPVVTFGQGTKLEIKR S307118G03 humanised L1
variable light (DNA sequence) SEQ ID 92
GACATCCAGATGACCCAGAGCCCCTCAAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACTATC
ACCTGCAGCGCCTCCCAGGGCATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCTAAGCTGCTGATCTACTACACCAGCAGCCTGCACAGCGGCGTGCCCAGCAGGTTCTCC
GGCAGCGGCAGCGGAACCGACTACACCCTGACCATTAGCAGCCTCCAGCCCGAGGACTTCGCC
ACCTACTACTGCCAGCAGTACAGCAAGCTGCCCTGGACCTTCGGCCAGGGCACCAAACTGGAG
ATCAAGCGT S307118G03 CDRH1 SEQ ID 93 DYYMK S307118G03 CDRH2 SEQ ID
94 EIYPNNGGITYNQKFKG S307118G03 CDRH3 SEQ ID 95 GYEFVY S307118G03
CDRL1 SEQ ID 96 SASQGISNYLN S307118G03 CDRL2 SEQ ID 97 YTSSLHS
S307118G03 CDRL3 SEQ ID 98 QQYSKLPWT S307118G03 humanised H5 CDRH3
SEQ ID 99 GYEFDY S307118G03 humanised H0 heavy chain SEQ ID 100
QVQLVQSGAEVKKPGSSVKVSCKASGGTFSDYYMKVVVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCARGYEFVYWGQGTLVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH
EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK
TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 humanised
H0 heavy chain (polynucleotide) SEQ ID 101
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCGGCACCTTCAGCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAGGGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S307118G03 humanised H1 heavy chain SEQ ID 102
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCARGYEFVYWGQGTLVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH
EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK
TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 humanised
H1 heavy chain (DNA sequence) SEQ ID 103
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAGGGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S307118G03 humanised H2 heavy chain SEQ ID 104
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCANGYEFVYWGQGTLVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH
EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK
TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 humanised
H2 heavy chain (DNA sequence) SEQ ID 105
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAACGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S307118G03 humanised H3 heavy chain SEQ ID 106
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWIGEIYPNNGGITYNQKFKG
RATLTVDKSTSTAYMELSSLRSEDTAVYYCANGYEFVYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
NHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE
DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI
SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG
SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 humanised
H3 heavy chain (DNA sequence) SEQ ID 107
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATAGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGCGACCCTCACCGTCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAACGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S307118G03 humanised H4 heavy chain SEQ ID 108
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWMGEIYPNNGGITYNQKFK
GRVTITADKSTSTAYMELSSLRSEDTAVYYCADGYEFVYWGQGTLVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN
VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH
EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK
TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSD
GSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 humanised
H4 heavy chain (DNA sequence) SEQ ID 109
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATGGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGTGACCATCACCGCCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCGACGGCTACGAGTTCGTGTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S307118G03 humanised H5 heavy chain SEQ ID 110
QVQLVQSGAEVKKPGSSVKVSCKASGYTFTDYYMKVVVRQAPGQGLEWIGEIYPNNGGITYNQKFKG
RATLTVDKSTSTAYMELSSLRSEDTAVYYCANGYEFDYWGQGTLVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV
NHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE
DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI
SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG
SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S307118G03 humanised
H5 heavy chain (DNA sequence) SEQ ID 111
CAGGTGCAGCTGGTGCAGAGCGGCGCCGAAGTGAAGAAGCCCGGCTCCAGCGTGAAGGTGAG
CTGCAAGGCTAGCGGCTACACCTTCACCGACTACTACATGAAGTGGGTGAGGCAGGCCCCCGG
CCAGGGACTGGAGTGGATAGGCGAGATCTACCCCAACAACGGGGGCATCACCTACAACCAGAA
GTTCAAGGGCAGGGCGACCCTCACCGTCGACAAAAGCACCAGCACCGCCTACATGGAACTGAG
CAGCCTGAGGAGCGAGGACACCGCCGTGTACTACTGCGCCAACGGCTACGAGTTCGACTATTG
GGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S307118G03 humanised L0 light chain SEQ ID 112
DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGS
GTDFTLTISSLQPEDFATYYCQQYSKLPVVTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL
LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC S307118G03 humanised L0 light chain (DNA sequence) SEQ
ID 113
GACATCCAGATGACCCAGAGCCCCTCAAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACTATC
ACCTGCAGCGCCTCCCAGGGCATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCTAAGCTGCTGATCTACTACACCAGCAGCCTGCACAGCGGCGTGCCCAGCAGGTTCTCC
GGCAGCGGCAGCGGAACCGACTTCACCCTGACCATTAGCAGCCTCCAGCCCGAGGACTTCGCC
ACCTACTACTGCCAGCAGTACAGCAAGCTGCCCTGGACCTTCGGCCAGGGCACCAAACTGGAG
ATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAG
AGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAG
TGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAG
CAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCA
CAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAA
CCGGGGCGAGTGC S307118G03 humanised L1 light chain SEQ ID 114
DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAPKLLIYYTSSLHSGVPSRFSGSGS
GTDYTLTISSLQPEDFATYYCQQYSKLPVVTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL
NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC S307118G03 humanised L1 light chain (DNA sequence) SEQ
ID 115
GACATCCAGATGACCCAGAGCCCCTCAAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACTATC
ACCTGCAGCGCCTCCCAGGGCATCAGCAACTACCTGAACTGGTACCAGCAGAAGCCCGGCAAG
GCCCCTAAGCTGCTGATCTACTACACCAGCAGCCTGCACAGCGGCGTGCCCAGCAGGTTCTCC
GGCAGCGGCAGCGGAACCGACTACACCCTGACCATTAGCAGCCTCCAGCCCGAGGACTTCGCC
ACCTACTACTGCCAGCAGTACAGCAAGCTGCCCTGGACCTTCGGCCAGGGCACCAAACTGGAG
ATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAG
AGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAG
TGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAG
CAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCA
CAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAA
CCGGGGCGAGTGC S332121F02 murine variable heavy chain SEQ ID 116
EVQLQQSGPVLVKPGASVKMSCEASGYTFTDYYMNVVVKQSHGKTLEWIGVINPYNGGTDYNQKFK
GKATLTVDKSSSTAYMELNSLTSEDSAVYYCARSVYDYPFDYWGQGTLVTVSS S332121F02
murine variable heavy chain (DNA sequence) SEQ ID 117
GAGGTGCAGCTGCAGCAGAGCGGCCCCGTGCTGGTGAAGCCTGGAGCCAGCGTGAAAATGAG
CTGCGAAGCCAGCGGCTACACCTTCACCGACTACTACATGAACTGGGTGAAGCAGAGCCACGG
CAAGACCCTGGAGTGGATCGGCGTGATCAACCCCTACAACGGGGGCACCGACTACAACCAGAA
GTTCAAGGGCAAGGCCACTCTGACCGTGGACAAGAGCTCCAGCACCGCCTACATGGAACTGAA
CAGCCTCACCTCTGAGGACAGCGCCGTCTATTACTGCGCCAGGAGCGTGTACGACTACCCCTTC
GACTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC S332121F02 chimeric heavy
chain SEQ ID 118
EVQLQQSGPVLVKPGASVKMSCEASGYTFTDYYMNVVVKQSHGKTLEWIGVINPYNGGTDYNQKFK
GKATLTVDKSSSTAYMELNSLTSEDSAVYYCARSVYDYPFDYWGQGTLVTVSSASTKGPSVFPLAPS
SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV
SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S332121F02
chimeric heavy chain (DNA sequence) SEQ ID 119
GAGGTGCAGCTGCAGCAGAGCGGCCCCGTGCTGGTGAAGCCTGGAGCCAGCGTGAAAATGAG
CTGCGAAGCCAGCGGCTACACCTTCACCGACTACTACATGAACTGGGTGAAGCAGAGCCACGG
CAAGACCCTGGAGTGGATCGGCGTGATCAACCCCTACAACGGGGGCACCGACTACAACCAGAA
GTTCAAGGGCAAGGCCACTCTGACCGTGGACAAGAGCTCCAGCACCGCCTACATGGAACTGAA
CAGCCTCACCTCTGAGGACAGCGCCGTCTATTACTGCGCCAGGAGCGTGTACGACTACCCCTTC
GACTACTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGT
GTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGG
TGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCG
TGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCG
TGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACAC
CAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCC
TGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCT
GATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGA
GGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGA
GGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCT
GAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACC
ATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGAT
GAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATC
GCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTG
GACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAG
GGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCC
TGAGCCTGTCCCCTGGCAAG
S332121F02 murine variable light chain SEQ ID 120
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
SGSETDFTLNIHPVEEEDAATYFCQQSIEDPRTFGGGTKLEIK S332121F02 murine
variable light chain (DNA sequence) SEQ ID 121
GACATCGTCCTGACCCAGAGCCCCGCCAGCCTGGCCGTGAGCCTGGGCCAGAGGGCCACAATC
AGCTGCAGGGCCTCTGAGTCCGTGAGCATCCACGGCACCCACCTGATGCACTGGTATCAGCAG
AAGCCCGGCCAGCCTCCCAAGCTGCTGATCTACGCCGCCAGCAACCTGGAGAGCGGCGTGCCC
GCTAGGTTCAGCGGAAGCGGCAGCGAGACCGACTTCACCCTGAACATCCACCCCGTGGAGGAG
GAAGACGCCGCCACCTACTTCTGCCAGCAGAGCATCGAGGACCCCAGGACCTTCGGCGGGGGC
ACCAAGCTCGAGATTAAGCGT S332121F02 chimeric light chain SEQ ID 122
MGWSCIILFLVATATGVHSDIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPK
LLIYAASNLESGVPARFSGSGSETDFTLNIHPVEEEDAATYFCQQSIEDPRTFGGGTKLEIKRTVAAP
VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL
SSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC S332121F02 chimeric light chain
(DNA sequence) SEQ ID 123
ATGGGCTGGTCCTGCATCATCCTGTTTCTGGTGGCCACCGCCACCGGCGTGCACAGCGACATC
GTCCTGACCCAGAGCCCCGCCAGCCTGGCCGTGAGCCTGGGCCAGAGGGCCACAATCAGCTG
CAGGGCCTCTGAGTCCGTGAGCATCCACGGCACCCACCTGATGCACTGGTATCAGCAGAAGCC
CGGCCAGCCTCCCAAGCTGCTGATCTACGCCGCCAGCAACCTGGAGAGCGGCGTGCCCGCTAG
GTTCAGCGGAAGCGGCAGCGAGACCGACTTCACCCTGAACATCCACCCCGTGGAGGAGGAAGA
CGCCGCCACCTACTTCTGCCAGCAGAGCATCGAGGACCCCAGGACCTTCGGCGGGGGCACCAA
GCTCGAGATTAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCA
GCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAA
GGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGC
AGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACG
AGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGA
GCTTCAACCGGGGCGAGTGC S322110D07 murine variable heavy chain SEQ ID
124
EVQLQQSGPELVKPGTSVKIPCKTSGYIFTDYSIDVVVKQSHGKSLEWIGDIDPNYGDPIYNHKFKGKA
TLTVDRSSSTAYMELRSLTSEDTAVYFCARRATGTDWFAFWGQGTLVTVSS S322110D07
murine variable heavy chain (DNA sequence) SEQ ID 125
GAGGTGCAGCTGCAGCAGAGCGGCCCCGAGCTGGTGAAACCCGGCACCAGCGTGAAGATCCC
CTGCAAGACCTCTGGCTACATCTTCACCGACTACAGCATCGACTGGGTGAAGCAGAGCCACGGC
AAGTCTCTGGAGTGGATTGGGGACATCGACCCCAACTACGGCGACCCCATCTACAACCACAAGT
TCAAGGGCAAGGCCACCCTGACCGTGGACAGGAGCAGCAGCACCGCCTACATGGAACTCAGGA
GCCTGACCAGCGAGGACACCGCCGTGTATTTTTGCGCCAGGAGGGCCACCGGCACTGATTGGT
TCGCCTTCTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC S322110D07 chimeric heavy
chain SEQ ID 126
EVQLQQSGPELVKPGTSVKIPCKTSGYIFTDYSIDVVVKQSHGKSLEWIGDIDPNYGDPIYNHKFKGKA
TLTVDRSSSTAYMELRSLTSEDTAVYFCARRATGTDWFAFWGQGTLVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS
HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S322110D07 chimeric
heavy chain (DNA sequence) SEQ ID 127
GAGGTGCAGCTGCAGCAGAGCGGCCCCGAGCTGGTGAAACCCGGCACCAGCGTGAAGATCCC
CTGCAAGACCTCTGGCTACATCTTCACCGACTACAGCATCGACTGGGTGAAGCAGAGCCACGGC
AAGTCTCTGGAGTGGATTGGGGACATCGACCCCAACTACGGCGACCCCATCTACAACCACAAGT
TCAAGGGCAAGGCCACCCTGACCGTGGACAGGAGCAGCAGCACCGCCTACATGGAACTCAGGA
GCCTGACCAGCGAGGACACCGCCGTGTATTTTTGCGCCAGGAGGGCCACCGGCACTGATTGGT
TCGCCTTCTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCG
TGTTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTG
GTGAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGC
GTGCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACC
GTGCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAAC
ACCAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGC
CCTGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACC
CTGATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCT
GAGGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGG
GAGGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGG
CTGAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAA
CCATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAG
ATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACAT
CGCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCT
GGACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCA
GGGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAG
CCTGAGCCTGTCCCCTGGCAAG S322110D07 murine variable light chain SEQ
ID 128
DIQMTQSPASLSVSVGETVTITCRASENIYNNLAWYQQKQGKSPQLLVYAATILADGVPSRFSGSGSG
TQYSLKINSLQSGDFGTYYCQHFWGTPLTFGAGTKLELKR S322110D07 murine variable
light chain (DNA sequence) SEQ ID 129
GACATCCAGATGACCCAGAGCCCCGCTAGCCTCAGCGTGTCCGTCGGCGAGACCGTGACCATC
ACCTGCAGGGCCAGCGAGAACATCTACAACAACCTGGCCTGGTATCAGCAGAAGCAGGGCAAA
AGCCCCCAGCTGCTGGTGTACGCCGCCACCATTCTGGCCGACGGCGTGCCCAGCAGGTTCTCT
GGAAGCGGCAGCGGCACCCAGTACAGCCTGAAGATCAACAGCCTGCAGAGCGGGGACTTCGG
CACCTACTACTGCCAGCACTTCTGGGGCACTCCCCTGACCTTCGGAGCCGGCACCAAGCTGGA
GCTGAAGCGT S322110D07 chimeric light chain SEQ ID 130
DIQMTQSPASLSVSVGETVTITCRASENIYNNLAWYQQKQGKSPQLLVYAATILADGVPSRFSGSGSG
TQYSLKINSLQSGDFGTYYCQHFWGTPLTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLL
NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLS
SPVTKSFNRGEC S322110D07 chimeric light chain (DNA sequence) SEQ ID
131 GACATCCAGATGACCCAGAGCCCCGCTAGCCTCAGCGTGTCCGTCGGCGAGACCGTGACCATC
ACCTGCAGGGCCAGCGAGAACATCTACAACAACCTGGCCTGGTATCAGCAGAAGCAGGGCAAA
AGCCCCCAGCTGCTGGTGTACGCCGCCACCATTCTGGCCGACGGCGTGCCCAGCAGGTTCTCT
GGAAGCGGCAGCGGCACCCAGTACAGCCTGAAGATCAACAGCCTGCAGAGCGGGGACTTCGG
CACCTACTACTGCCAGCACTTCTGGGGCACTCCCCTGACCTTCGGAGCCGGCACCAAGCTGGA
GCTGAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAA
GAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCA
GTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACA
GCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGC
ACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCA
ACCGGGGCGAGTGC S332126E04 murine variable heavy chain SEQ ID 132
QVQLQQPGAELVKPGASVKLSCKASGYTFTNYWMHVVVKQRPGQGLEWIGIIHPNSGSTNYNEKFKS
KATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGIYDYPFAYWGQGTLVTVSS S332126E04
murine variable heavy chain (DNA sequence) SEQ ID 133
CAGGTGCAGCTCCAGCAGCCCGGAGCCGAACTGGTGAAGCCCGGAGCCAGCGTCAAACTGTCC
TGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAAGCAGAGGCCCGGC
CAGGGCCTGGAGTGGATCGGCATCATCCACCCCAACAGCGGGAGCACCAACTACAACGAGAAG
TTCAAGAGCAAGGCCACCCTGACCGTGGACAAGAGCAGCAGCACTGCCTACATGCAGCTGAGC
AGCCTGACCAGCGAGGACAGCGCTGTGTACTACTGCGCCAGGGGCATCTACGACTACCCCTTC
GCCTATTGGGGCCAGGGCACACTAGTGACCGTGTCCAGC S332126E04 Chimeric heavy
chain SEQ ID 134
QVQLQQPGAELVKPGASVKLSCKASGYTFTNYWMHVVVKQRPGQGLEWIGIIHPNSGSTNYNEKFKS
KATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGIYDYPFAYWGQGTLVTVSSASTKGPSVFPLAPSS
KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV
SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S332126E04
Chimeric heavy chain (DNA sequence) SEQ ID 135
CAGGTGCAGCTCCAGCAGCCCGGAGCCGAACTGGTGAAGCCCGGAGCCAGCGTCAAACTGTCC
TGCAAGGCCAGCGGCTACACCTTCACCAACTACTGGATGCACTGGGTGAAGCAGAGGCCCGGC
CAGGGCCTGGAGTGGATCGGCATCATCCACCCCAACAGCGGGAGCACCAACTACAACGAGAAG
TTCAAGAGCAAGGCCACCCTGACCGTGGACAAGAGCAGCAGCACTGCCTACATGCAGCTGAGC
AGCCTGACCAGCGAGGACAGCGCTGTGTACTACTGCGCCAGGGGCATCTACGACTACCCCTTC
GCCTATTGGGGCCAGGGCACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTG
TTCCCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGT
GAAGGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGT
GCACACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGT
GCCCAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACAC
CAAGGTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCC
TGCCCCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCT
GATGATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGA
GGTGAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGA
GGAGCAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCT
GAACGGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACC
ATCAGCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGAT
GAGCTGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATC
GCCGTGGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTG
GACAGCGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAG
GGCAACGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCC
TGAGCCTGTCCCCTGGCAAG S332126E04 murine variable light chain SEQ ID
136
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
SGSETDFTLNIHPVEEEDAATYFCQQSIEDPYTFGGGTKLEIKR S332126E04 murine
variable light chain (DNA sequence) SEQ ID 137
GACATCGTGCTGACCCAGTCTCCCGCTAGCCTGGCCGTGTCTCTGGGCCAGAGGGCCACAATC
AGCTGCAGGGCCAGCGAGAGCGTCAGCATTCACGGCACCCACCTGATGCACTGGTACCAGCAG
AAGCCCGGCCAGCCTCCCAAGCTCCTGATCTACGCCGCCAGCAACCTGGAAAGCGGAGTGCCC
GCCAGGTTCAGCGGCAGCGGCTCCGAGACCGACTTCACCCTGAACATCCACCCCGTGGAGGAG
GAGGACGCCGCCACCTACTTCTGCCAGCAGAGCATCGAGGACCCCTACACCTTCGGCGGCGGC
ACCAAGCTGGAGATCAAGCGT S332126E04 Chimeric light chain SEQ ID 138
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
SGSETDFTLNIHPVEEEDAATYFCQQSIEDPYTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASV
VCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGEC S332126E04 Chimeric light chain (DNA sequence) SEQ
ID 139
GACATCGTGCTGACCCAGTCTCCCGCTAGCCTGGCCGTGTCTCTGGGCCAGAGGGCCACAATC
AGCTGCAGGGCCAGCGAGAGCGTCAGCATTCACGGCACCCACCTGATGCACTGGTACCAGCAG
AAGCCCGGCCAGCCTCCCAAGCTCCTGATCTACGCCGCCAGCAACCTGGAAAGCGGAGTGCCC
GCCAGGTTCAGCGGCAGCGGCTCCGAGACCGACTTCACCCTGAACATCCACCCCGTGGAGGAG
GAGGACGCCGCCACCTACTTCTGCCAGCAGAGCATCGAGGACCCCTACACCTTCGGCGGCGGC
ACCAAGCTGGAGATCAAGCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGAT
GAGCAGCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAG
GCCAAGGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGAC
CGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGA
CTACGAGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGAC
CAAGAGCTTCAACCGGGGCGAGTGC S336105A07 murine variable heavy chain
SEQ ID 140
EVKLLQSGGGLVQPGGSLKLSCAASGIDFSRYWMSWVRRAPGKGLEWIGEINPDRSTINYAPSLKDK
FIISRDNAKNTLYLQMSKVRSEDTALYYCAVFYYDYEGAMDYWGQGTSVTVSS S336105A07
murine variable heavy chain (DNA sequence) SEQ ID 141
GAGGTGAAGCTTCTCCAGTCTGGAGGTGGCCTGGTGCAGCCTGGAGGATCCCTGAAACTCTCCT
GTGCAGCCTCAGGAATCGATTTTAGTAGATACTGGATGAGTTGGGTTCGGCGGGCTCCAGGGAA
AGGACTAGAATGGATTGGAGAAATTAATCCAGATAGGAGTACAATCAACTATGCACCATCTCTAA
AGGATAAATTCATCATCTCCAGAGACAACGCCAAAAATACGCTGTACCTGCAAATGAGCAAAGTG
AGATCTGAGGACACAGCCCTTTATTACTGTGCAGTTTTCTACTATGATTACGAGGGTGCTATGGA
CTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA S336105A07 Chimeric heavy
chain SEQ ID 142
EVKLLQSGGGLVQPGGSLKLSCAASGIDFSRYWMSWVRRAPGKGLEWIGEINPDRSTINYAPSLKDK
FIISRDNAKNTLYLQMSKVRSEDTALYYCAVFYYDYEGAMDYWGQGTSVTVSSAKTTAPSVFPLAPS
SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI
CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV
SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S336105A07
Chimeric heavy chain (DNA sequence) SEQ ID 143
GAGGTGAAGCTTCTCCAGTCTGGAGGTGGCCTGGTGCAGCCTGGAGGATCCCTGAAACTCTCCT
GTGCAGCCTCAGGAATCGATTTTAGTAGATACTGGATGAGTTGGGTTCGGCGGGCTCCAGGGAA
AGGACTAGAATGGATTGGAGAAATTAATCCAGATAGGAGTACAATCAACTATGCACCATCTCTAA
AGGATAAATTCATCATCTCCAGAGACAACGCCAAAAATACGCTGTACCTGCAAATGAGCAAAGTG
AGATCTGAGGACACAGCCCTTTATTACTGTGCAGTTTTCTACTATGATTACGAGGGTGCTATGGA
CTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCAGCCAAAACAACAGCCCCCAGCGTGTTC
CCCCTGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAA
GGACTACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCA
CACCTTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCC
CAGCAGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAG
GTGGACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCC
CCCGAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATG
ATCAGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTG
AAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAG
CAGTACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAAC
GGCAAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCA
GCAAGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGC
TGACCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGT
GGAGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAG
CGATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAA
CGTGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGC
CTGTCCCCTGGCAAG S336105A07 murine varaible light chain SEQ ID 144
DIVMTQSQKFMSTSVGDRVSVTCKASQNVDTNVAWYQQKPGQSPKALIYSASYRFSGVPDRFTGSG
SGTDFTLTISNVQSEDLAEYFCQQYNSFPFTFGSGTKLEIKR S336105A07 murine
variable light chain (DNA sequence) SEQ ID 145
GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCAC
CTGCAAGGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAAAAACCAGGGCAATCTC
CTAAAGCACTGATTTACTCGGCATCCTACCGGTTCAGTGGAGTCCCTGATCGCTTCACAGGCAGT
GGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTATTT
CTGTCAGCAATATAACAGCTTTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAACGT
S336105A07 chimeric light chain SEQ ID 146
DIVMTQSQKFMSTSVGDRVSVTCKASQNVDTNVAWYQQKPGQSPKALIYSASYRFSGVPDRFTGSG
SGTDFTLTISNVQSEDLAEYFCQQYNSFPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVC
LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL
SSPVTKSFNRGEC S336105A07 chimeric light chain (DNA sequence) SEQ ID
147
GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACATCAGTAGGAGACAGGGTCAGCGTCAC
CTGCAAGGCCAGTCAGAATGTGGATACTAATGTAGCCTGGTATCAACAAAAACCAGGGCAATCTC
CTAAAGCACTGATTTACTCGGCATCCTACCGGTTCAGTGGAGTCCCTGATCGCTTCACAGGCAGT
GGATCTGGGACAGATTTCACTCTCACCATCAGCAATGTGCAGTCTGAAGACTTGGCAGAGTATTT
CTGTCAGCAATATAACAGCTTTCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAACGTA
CGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGCTGAAGAGCGGCACCG
CCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGGTGCAGTGGAAGGTGG
ACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAGGACAGCAAGGACTCCA
CCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACG
CCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGT GC
S335115G01 murine variable heavy chain SEQ ID 148
PVQLQQPGTELVRPGTSVKLSCKASGYTFTSYWMHVVVKQRPGQGLEWIGVIDPSDSYTNYNQKFK
GKATLTVDTSSSTAYMQLSSLTSEDSAVYYCARQVFDYPMDYWGQGTSVTVSS S335115G01
murine variable heavy chain (DNA sequence) SEQ ID 149
CCGGTCCAACTGCAGCAGCCTGGGACTGAGCTGGTGAGGCCTGGGACTTCAGTGAAGTTGTCC
TGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTAAAGCAGAGGCCTGGAC
AAGGCCTTGAGTGGATCGGAGTGATTGATCCTTCTGATAGTTATACTAACTACAATCAAAAGTTCA
AGGGCAAGGCCACATTGACTGTAGACACATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCT
GACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGACAGGTGTTTGACTATCCTATGGACTACT
GGGGTCAAGGAACCTCAGTCACCGTCTCCTCA S335115G01 Chimeric heavy chain
SEQ ID 150
PVQLQQPGTELVRPGTSVKLSCKASGYTFTSHWMHVVVKQRPGQGLEWIGVIDPSDSYTNYNQKFK
GKATLTVDTSSSTAYMQLSSLTSEDSAVYYCARQVFDYPMDYWGQGTLVTVSSASTKGPSVFPLAP
SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT
YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD
VSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP
IEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S335115G01
Chimeric heavy chain (DNA sequence) SEQ ID 151
CCGGTCCAACTGCAGCAGCCTGGGACTGAGCTGGTGAGGCCTGGGACTTCAGTGAAGTTGTCC
TGCAAGGCTTCTGGCTACACCTTCACCAGCCACTGGATGCACTGGGTAAAGCAGAGGCCTGGAC
AAGGCCTTGAGTGGATCGGAGTGATTGATCCTTCTGATAGTTATACTAACTACAATCAAAAGTTCA
AGGGCAAGGCCACATTGACTGTAGACACATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCT
GACATCTGAGGACTCTGCGGTCTATTACTGTGCAAGACAGGTGTTTGACTATCCTATGGACTACT
GGGGTCAAGGAACACTAGTGACCGTGTCCAGCGCCAGCACCAAGGGCCCCAGCGTGTTCCCCC
TGGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGAC
TACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACC
TTCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGC
AGCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTG
GACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCC
GAGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATC
AGCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAG
TTCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAG
TACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGC
AAGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCA
AGGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGA
CCAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGG
AGTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCG
ATGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACG
TGTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCT
GTCCCCTGGCAAG S335115G01 murine variable light chain SEQ ID 152
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
SGSETDFTLNIHPVEEEDAATYFCQQSIEDPVVTFGGGTKLEIKR S335115G01 murine
variable light chain (DNA sequence) SEQ ID 153
GACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCT
CCTGCAGAGCCAGTGAAAGTGTCAGTATTCATGGTACTCATTTAATGCACTGGTACCAACAGAAA
CCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACCTAGAATCTGGAGTCCCTGCCA
GGTTCAGTGGCAGTGGGTCTGAGACAGACTTCACCCTCAACATCCATCCTGTGGAGGAGGAGGA
TGCTGCAACCTATTTCTGTCAGCAAAGTATTGAGGATCCGTGGACGTTCGGTGGAGGCACCAAG
CTGGAAATCAAACGT S335115G01 Chimeric light chain SEQ ID 154
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
SGSETDFTLNIHPVEEEDAATYFCQQSIEDPVVTFGGGTKLEINRTVAAPSVFIFPPSDEQLKSGTAS
VVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGEC S335115G01 Chimeric light chain (DNA sequence) SEQ
ID 155
GACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCT
CCTGCAGAGCCAGTGAAAGTGTCAGTATTCATGGTACTCATTTAATGCACTGGTACCAACAGAAA
CCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACCTAGAATCTGGAGTCCCTGCCA
GGTTCAGTGGCAGTGGGTCTGAGACAGACTTCACCCTCAACATCCATCCTGTGGAGGAGGAGGA
TGCTGCAACCTATTTCTGTCAGCAAAGTATTGAGGATCCGTGGACGTTCGGTGGAGGCACCAAG
CTGGAAATCAATCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCAGC
TGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAAGG
TGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGCAG
GACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAG
AAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGC
TTCAACCGGGGCGAGTGC S335122F05 murine variable heavy chain SEQ ID
156
QVQLQQSGAELVRPGASVTLSCKASGYTFTDYEMHVVVKQTPVHGLEWIGAIDPETGGTAYNQKFKG
KAILTADKSSSTAYMELRSLTSEDSAVYYCTRSIYDYYFDYWGQGTTLTVSS S335122F05
murine variable heavy chain (DNA sequence) SEQ ID 157
CAGGTTCAACTGCAGCAGTCTGGGGCTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCC
TGCAAGGCTTCGGGCTACACATTTACTGACTATGAAATGCACTGGGTGAAGCAGACACCTGTGC
ATGGCCTGGAATGGATTGGAGCTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTC
AAGGGCAAGGCCATACTGACTGCAGACAAATCCTCCAGCACAGCCTACATGGAGCTCCGCAGCC
TGACATCTGAGGACTCTGCCGTCTATTACTGTACAAGATCGATTTATGATTACTACTTTGACTACT
GGGGCCAAGGCACCACTCTCACAGTCTCCTCA S335122F05 Chimeric heavy chain
SEQ ID 158
QVQLQQSGAELVRPGASVTLSCKASGYTFTDYEMHVVVKQTPVHGLEWIGAIDPETGGTAYNQKFKG
KAILTADKSSSTAYMELRSLTSEDSAVYYCTRSIYDYYFDYWGQGTTLTVSSAKTTPPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC
NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS
HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE
KTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS
DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK S335122F05 Chimeric
heavy chain (DNA sequence) SEQ ID 159
CAGGTTCAACTGCAGCAGTCTGGGGCTGAGCTGGTGAGGCCTGGGGCTTCAGTGACGCTGTCC
TGCAAGGCTTCGGGCTACACATTTACTGACTATGAAATGCACTGGGTGAAGCAGACACCTGTGC
ATGGCCTGGAATGGATTGGAGCTATTGATCCTGAAACTGGTGGTACTGCCTACAATCAGAAGTTC
AAGGGCAAGGCCATACTGACTGCAGACAAATCCTCCAGCACAGCCTACATGGAGCTCCGCAGCC
TGACATCTGAGGACTCTGCCGTCTATTACTGTACAAGATCGATTTATGATTACTACTTTGACTACT
GGGGCCAAGGCACCACTCTCACAGTCTCCTCAGCCAAAACGACACCCCCCAGCGTGTTCCCCCT
GGCCCCCAGCAGCAAGAGCACCAGCGGCGGCACAGCCGCCCTGGGCTGCCTGGTGAAGGACT
ACTTCCCCGAACCGGTGACCGTGTCCTGGAACAGCGGAGCCCTGACCAGCGGCGTGCACACCT
TCCCCGCCGTGCTGCAGAGCAGCGGCCTGTACAGCCTGAGCAGCGTGGTGACCGTGCCCAGCA
GCAGCCTGGGCACCCAGACCTACATCTGTAACGTGAACCACAAGCCCAGCAACACCAAGGTGG
ACAAGAAGGTGGAGCCCAAGAGCTGTGACAAGACCCACACCTGCCCCCCCTGCCCTGCCCCCG
AGCTGCTGGGAGGCCCCAGCGTGTTCCTGTTCCCCCCCAAGCCTAAGGACACCCTGATGATCA
GCAGAACCCCCGAGGTGACCTGTGTGGTGGTGGATGTGAGCCACGAGGACCCTGAGGTGAAGT
TCAACTGGTACGTGGACGGCGTGGAGGTGCACAATGCCAAGACCAAGCCCAGGGAGGAGCAGT
ACAACAGCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCA
AGGAGTACAAGTGTAAGGTGTCCAACAAGGCCCTGCCTGCCCCTATCGAGAAAACCATCAGCAA
GGCCAAGGGCCAGCCCAGAGAGCCCCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGAC
CAAGAACCAGGTGTCCCTGACCTGCCTGGTGAAGGGCTTCTACCCCAGCGACATCGCCGTGGA
GTGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGA
TGGCAGCTTCTTCCTGTACAGCAAGCTGACCGTGGACAAGAGCAGATGGCAGCAGGGCAACGT
GTTCAGCTGCTCCGTGATGCACGAGGCCCTGCACAATCACTACACCCAGAAGAGCCTGAGCCTG
TCCCCTGGCAAG S335122F05 murine variable light chain SEQ ID 160
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
GGSETDFTLNIHPVEEEDGATYFCQQSIEYPRTFGGGTKLEINR S335122F05 murine
variable light chain (DNA sequence) SEQ ID 161
GACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCT
CCTGCAGAGCCAGTGAAAGTGTCAGTATTCATGGTACTCATTTAATGCACTGGTACCAACAGAAA
CCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACCTAGAATCTGGAGTCCCTGCCA
GGTTCAGTGGCGGTGGGTCTGAGACAGACTTCACCCTCAACATCCATCCTGTGGAGGAGGAGG
ATGGTGCAACCTATTTCTGTCAGCAAAGTATTGAGTATCCTCGGACGTTCGGTGGAGGCACCAA
GCTGGAAATCAATCGT S335122F05 Chimeric light chain SEQ ID 162
DIVLTQSPASLAVSLGQRATISCRASESVSIHGTHLMHWYQQKPGQPPKLLIYAASNLESGVPARFSG
GGSETDFTLNIHPVEEEDGATYFCQQSIEYPRTFGGGTKLEINRTVAAPSVFIFPPSDEQLKSGTASVV
CLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ
GLSSPVTKSFNRGEC S335122F05 Chimeric light chain (DNA sequence) SEQ
ID 163
GACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCT
CCTGCAGAGCCAGTGAAAGTGTCAGTATTCATGGTACTCATTTAATGCACTGGTACCAACAGAAA
CCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACCTAGAATCTGGAGTCCCTGCCA
GGTTCAGTGGCGGTGGGTCTGAGACAGACTTCACCCTCAACATCCATCCTGTGGAGGAGGAGG
ATGGTGCAACCTATTTCTGTCAGCAAAGTATTGAGTATCCTCGGACGTTCGGTGGAGGCACCAA
GCTGGAAATCAATCGTACGGTGGCCGCCCCCAGCGTGTTCATCTTCCCCCCCAGCGATGAGCA
GCTGAAGAGCGGCACCGCCAGCGTGGTGTGTCTGCTGAACAACTTCTACCCCCGGGAGGCCAA
GGTGCAGTGGAAGGTGGACAATGCCCTGCAGAGCGGCAACAGCCAGGAGAGCGTGACCGAGC
AGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACG
AGAAGCACAAGGTGTACGCCTGTGAGGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGA
GCTTCAACCGGGGCGAGTGC S332121F02 CDRH1 SEQ.I.D.NO: 164 DYYNM
S332121F02 CDRH2 SEQ.I.D.NO: 165 VINPYNGGTDYNQKFG S332121F02 CDRH3
SEQ.I.D.NO: 166 SVYDYPFDY S332121 F02 CDRL1 SEQ.I.D.NO: 167
RASESVSIHGTHLMH S332121 F02 CDRL2 SEQ.I.D.NO: 168 AASNLES S332121
F02 CDRL3 SEQ.I.D.NO: 169 QQSIEDPRT S322110D07 CDRH1 SEQ.I.D.NO:
170 DYSID S322110D07 CDRH2 SEQ.I.D.NO: 171 DIDPNYGDPIYNHKFKG
S322110D07 CDRH3 SEQ.I.D.NO: 172 RATGTDWFAF S322110D07 CDRL1
SEQ.I.D.NO: 173 RASENIYNNLA S322110D07 CDRL2 SEQ.I.D.NO: 174
AATILAD S322110D07 CDRL3 SEQ.I.D.NO: 175
QHFWGTPLT S332126E04 CDRH1 SEQ.I.D.NO: 176 NYWMH S332126E04 CDRH2
SEQ.I.D.NO: 177 IIHPNSGSTNYNEKFKS S332126E04 CDRH3 SEQ.I.D.NO: 178
GIYDYPFAY S332126E04 CDRL1 SEQ.I.D.NO: 179 RASESVSIHGTHLMH
S332126E04 CDRL2 SEQ.I.D.NO: 180 AASNLES S332126E04 CDRL3
SEQ.I.D.NO: 181 QQSIEDPYT S336105A07 CDRH1 SEQ.I.D.NO: 182 RYWMS
S336105A07 CDRH2 SEQ.I.D.NO: 183 EINPDRSTINYAPSLKD S336105A07 CDRH3
SEQ.I.D.NO: 184 FYYDYEGAMDY S336105A07 CDRL1 SEQ.I.D.NO: 185
KASQNVDTNVA S336105A07 CDRL2 SEQ.I.D.NO: 186 SASYRFS S336105A07
CDRL3 SEQ.I.D.NO: 187 QQYNSFPFT S335115G01 CDRH1 SEQ.I.D.NO: 188
SYWMH S335115G01 CDRH2 SEQ.I.D.NO: 189 VIDPSDSYTNYNQKFKG S335115G01
CDRH3 SEQ.I.D.NO: 190 QVFDYPMDY S335115G01 CDRL1 SEQ.I.D.NO: 191
RASESVSIHGTHLMH S335115G01 CDRL2 SEQ.I.D.NO: 192 AASNLES S335115G01
CDRL3 SEQ.I.D.NO: 193 QQSIEDPVVT S335122F05 CDRH1 SEQ.I.D.NO: 194
DYEMH S335122F05 CDRH2 SEQ.I.D.NO: 195 AIDPETGGTAYNQKFKG S335122F05
CDRH3 SEQ.I.D.NO: 196 SIYDYYFDY S335122F05 CDRL1 SEQ.I.D.NO: 197
RASESVSIHGTHLMH S335122F05 CDRL2 SEQ.I.D.NO: 198 AASNLES S335122F05
CDRL3 SEQ.I.D.NO: 199 QQSIEYPRT
Sequence CWU 1
1
19915PRTmus musculus 1Asn Tyr Trp Met His1 5 217PRTmus musculus
2Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe Lys1 5
10 15 Gly312PRTmus musculus 3Gly Ala Ile Tyr Asn Gly Tyr Asp Val
Leu Asp Asn1 5 10 411PRTmus musculus 4Ser Ala Ser Gln Asp Ile Ser
Asn Tyr Leu Asn1 5 10 57PRTmus musculus 5Tyr Thr Ser Asn Leu His
Ser1 5 69PRTmus musculus 6Gln Gln Tyr Arg Lys Leu Pro Trp Thr1 5
7121PRTmus musculus 7Glu Val Gln Leu Gln Gln Ser Gly Ala Val Leu
Ala Arg Pro Gly Ala1 5 10 15 Ser Val Lys Met Ser Cys Lys Gly Ser
Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Lys Gln
Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ala Thr Tyr Arg
Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys
Ala Lys Leu Thr Ala Val Thr Ser Thr Ser Thr Ala Tyr65 70 75 80 Met
Glu Leu Ser Ser Leu Thr Asn Glu Asp Ser Ala Val Tyr Tyr Cys 85 90
95 Thr Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val Leu Asp Asn Trp Gly
100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 8363DNAmus
musculus 8gaggtgcagc tgcagcagag cggcgccgtg ctggccaggc ccggagctag
cgtgaagatg 60agctgcaagg gcagcggcta caccttcacc aactactgga tgcactgggt
gaaacagagg 120cccggccagg gactggagtg gatcggcgcc acctacaggg
gccacagcga cacctactac 180aaccagaagt tcaagggcaa ggccaagctg
accgccgtga cctcaaccag caccgcctac 240atggaactga gcagcctgac
caacgaggac agcgccgtct attactgcac caggggcgcc 300atctacaacg
gctacgacgt gctggacaat tggggccagg gaacactagt gaccgtgtcc 360agc
3639108PRTmus musculus 9Asp Ile Gln Leu Thr Gln Thr Thr Ser Ser Leu
Ser Ala Ser Leu Gly1 5 10 15 Asp Arg Val Thr Ile Ser Cys Ser Ala
Ser Gln Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys
Pro Asp Gly Thr Val Glu Leu Val Ile 35 40 45 Tyr Tyr Thr Ser Asn
Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser
Gly Thr Asp Tyr Ser Leu Thr Ile Gly Tyr Leu Glu Pro65 70 75 80 Glu
Asp Val Ala Thr Tyr Tyr Cys Gln Gln Tyr Arg Lys Leu Pro Trp 85 90
95 Thr Phe Gly Gly Gly Ser Lys Leu Glu Ile Lys Arg 100 105
10324DNAmus musculus 10gatatccagc tgacccagac cacaagcagc ctgagcgcct
ccctgggcga cagggtgacc 60attagctgca gcgccagcca ggacatcagc aactacctga
actggtacca gcagaagccc 120gacggcaccg tggagctcgt gatctactac
acctccaacc tgcacagcgg cgtgcccagc 180aggttctctg gcagcggcag
cggcaccgac tacagcctga ccatcggcta tctggagccc 240gaggacgtcg
ccacctacta ctgccagcag tacaggaagc tgccctggac cttcggcgga
300ggctctaagc tggagattaa gcgt 32411121PRTArtificial
SequenceHumansed antibody sequence 11Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 12363DNAArtificial SequenceHumansed antibody sequence
12caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg ccagcggcgg caccttcagc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
agagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcgc caggggcgcc 300atctacaacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agc
36313121PRTArtificial SequenceHumansed antibody sequence 13Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20
25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Ala Ile Tyr Asn
Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val
Thr Val Ser Ser 115 120 14363DNAArtificial SequenceHumansed
antibody sequence 14caggtgcagc tggtccagag cggcgccgaa gtgaagaagc
ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta caccttcacc aactactgga
tgcactgggt gaggcaggcc 120cccggacagg gcctggagtg gatgggcgcc
acctacaggg gccacagcga cacctactac 180aaccagaagt tcaagggccg
ggtgaccatc accgccgaca agagcaccag caccgcctac 240atggaactga
gcagcctcag gagcgaggac accgctgtgt attactgcgc caggggcgcc
300atctacaacg gctacgacgt gctggacaac tggggccagg gcacactagt
gaccgtgtcc 360agc 36315121PRTArtificial SequenceHumansed antibody
sequence 15Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro
Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser
Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile
Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser
Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg
Gly Ala Ile Tyr Asn Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 16363DNAArtificial
SequenceHumansed antibody sequence 16caggtgcagc tggtccagag
cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta
caccttcacc aactactgga tgcactgggt gaggcaggcc 120cccggacagg
gcctggagtg gatgggcgcc acctacaggg gccacagcga cacctactac
180aaccagaagt tcaagggccg ggtgaccatc accgccgaca agagcaccag
caccgcctac 240atggaactga gcagcctcag gagcgaggac accgctgtgt
attactgcac caggggcgcc 300atctacaacg gctacgacgt gctggacaac
tggggccagg gcacactagt gaccgtgtcc 360agc 36317121PRTArtificial
SequenceHumansed antibody sequence 17Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 18363DNAArtificial SequenceHumansed antibody sequence
18caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg gcagcggcta caccttcacc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
cgagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcac caggggcgcc 300atctacaacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agc
36319121PRTArtificial SequenceHumansed antibody sequence 19Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asn Tyr 20
25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Ile 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Thr Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asn
Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val
Thr Val Ser Ser 115 120 20363DNAArtificial SequenceHumansed
antibody sequence 20caggtgcagc tggtccagag cggcgccgaa gtgaagaagc
ccggcagctc cgtgaaagtg 60agctgcaagg gcagcggcta caccttcacc aactactgga
tgcactgggt gaggcaggcc 120cccggacagg gcctggagtg gatcggcgcc
acctacaggg gccacagcga cacctactac 180aaccagaagt tcaagggccg
ggcgaccctc accgccgaca cgagcaccag caccgcctac 240atggaactga
gcagcctcag gagcgaggac accgctgtgt attactgcac caggggcgcc
300atctacaacg gctacgacgt gctggacaac tggggccagg gcacactagt
gaccgtgtcc 360agc 36321121PRTArtificial SequenceHumansed antibody
sequence 21Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro
Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr
Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser
Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile
Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser
Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg
Gly Ala Ile Tyr Asp Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 22363DNAArtificial
SequenceHumansed antibody sequence 22caggtgcagc tggtccagag
cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg gcagcggcta
caccttcacc aactactgga tgcactgggt gaggcaggcc 120cccggacagg
gcctggagtg gatgggcgcc acctacaggg gccacagcga cacctactac
180aaccagaagt tcaagggccg ggtgaccatc accgccgaca cgagcaccag
caccgcctac 240atggaactga gcagcctcag gagcgaggac accgctgtgt
attactgcac caggggcgcc 300atctacgacg gctacgacgt gctggacaac
tggggccagg gcacactagt gaccgtgtcc 360agc 36323121PRTArtificial
SequenceHumansed antibody sequence 23Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Gly Ala Ile Tyr Asp Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 24363DNAArtificial SequenceHumansed antibody sequence
24caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg ccagcggcgg caccttcagc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
agagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcgc caggggcgcc 300atctacgacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agc
36325121PRTArtificial SequenceHumansed antibody sequence 25Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20
25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Ala Ile Tyr Asp
Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val
Thr Val Ser Ser 115 120 26363DNAArtificial SequenceHumansed
antibody sequence 26caggtgcagc tggtccagag cggcgccgaa gtgaagaagc
ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta caccttcacc aactactgga
tgcactgggt gaggcaggcc 120cccggacagg gcctggagtg gatgggcgcc
acctacaggg gccacagcga cacctactac 180aaccagaagt tcaagggccg
ggtgaccatc accgccgaca agagcaccag caccgcctac 240atggaactga
gcagcctcag gagcgaggac accgctgtgt attactgcgc caggggcgcc
300atctacgacg gctacgacgt gctggacaac tggggccagg gcacactagt
gaccgtgtcc 360agc 36327121PRTArtificial SequenceHumansed antibody
sequence 27Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro
Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly
Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser
Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile
Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser
Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg
Gly Ala Ile Tyr Asp Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110
Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 28363DNAArtificial
SequenceHumansed antibody sequence 28caggtgcagc tggtccagag
cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta
caccttcacc aactactgga tgcactgggt gaggcaggcc 120cccggacagg
gcctggagtg gatgggcgcc acctacaggg gccacagcga cacctactac
180aaccagaagt tcaagggccg ggtgaccatc accgccgaca agagcaccag
caccgcctac 240atggaactga gcagcctcag gagcgaggac accgctgtgt
attactgcac caggggcgcc 300atctacgacg gctacgacgt gctggacaac
tggggccagg gcacactagt gaccgtgtcc 360agc 36329121PRTArtificial
SequenceHumansed antibody sequence 29Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45
Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50
55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asp Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 30363DNAArtificial SequenceHumansed antibody sequence
30caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg gcagcggcta caccttcacc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatcggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggcgaccctc accgccgaca
cgagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcac caggggcgcc 300atctacgacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agc
36331108PRTArtificial SequenceHumansed antibody sequence 31Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Asp Ile Ser Asn Tyr 20
25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile 35 40 45 Tyr Tyr Thr Ser Asn Leu His Ser Gly Val Pro Ser Arg
Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Ser Leu Gln Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Tyr Arg Lys Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys
Leu Glu Ile Lys Arg 100 105 32324DNAArtificial SequenceHumansed
antibody sequence 32gacatccaga tgacccagag ccctagctca ctgagcgcca
gcgtgggcga cagggtgacc 60attacctgct ccgccagcca ggacatcagc aactacctga
actggtacca gcagaagccc 120ggcaaggccc ccaagctgct gatctactac
acctccaacc tgcactccgg cgtgcccagc 180aggttcagcg gaagcggcag
cggcaccgat ttcaccctga ccatctccag cctgcagccc 240gaggacttcg
ccacctacta ctgccagcag tacaggaagc tcccctggac tttcggccag
300ggcaccaaac tggagatcaa gcgt 32433108PRTArtificial
SequenceHumansed antibody sequence 33Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15 Asp Arg Val Thr Ile
Thr Cys Ser Ala Ser Gln Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp
Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr
Tyr Thr Ser Asn Leu His Ser 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
Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Arg Lys
Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
Arg 100 105 34324DNAArtificial SequenceHumansed antibody sequence
34gacatccaga tgacccagag ccctagctca ctgagcgcca gcgtgggcga cagggtgacc
60attacctgct ccgccagcca ggacatcagc aactacctga actggtacca gcagaagccc
120ggcaaggccc ccaagctgct gatctactac acctccaacc tgcactccgg
cgtgcccagc 180aggttcagcg gaagcggcag cggcaccgat tacaccctga
ccatctccag cctgcagccc 240gaggacttcg ccacctacta ctgccagcag
tacaggaagc tcccctggac tttcggccag 300ggcaccaaac tggagatcaa gcgt
32435108PRTArtificial SequenceHumansed antibody sequence 35Asp Ile
Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Asp Ile Ser Asn Tyr 20
25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Glu Leu Val
Ile 35 40 45 Tyr Tyr Thr Ser Asn Leu His Ser 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 Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Tyr Arg Lys Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys
Leu Glu Ile Lys Arg 100 105 36324DNAArtificial SequenceHumansed
antibody sequence 36gacatccagc tgacccagag ccctagctca ctgagcgcca
gcgtgggcga cagggtgacc 60attacctgct ccgccagcca ggacatcagc aactacctga
actggtacca gcagaagccc 120ggcaaggccc ccgagctggt gatctactac
acctccaacc tgcactccgg cgtgcccagc 180aggttcagcg gaagcggcag
cggcaccgat tacaccctga ccatctccag cctgcagccc 240gaggacttcg
ccacctacta ctgccagcag tacaggaagc tcccctggac tttcggccag
300ggcaccaaac tggagatcaa gcgt 32437310PRThomo sapiens 37Met Pro Leu
Leu Leu Leu Leu Pro Leu Leu Trp Ala Gly Ala Leu Ala1 5 10 15 Met
Leu Gln Met Ala Gly Gln Cys Ser Gln Asn Glu Tyr Phe Asp Ser 20 25
30 Leu Leu His Ala Cys Ile Pro Cys Gln Leu Arg Cys Ser Ser Asn Thr
35 40 45 Pro Pro Leu Thr Cys Gln Arg Tyr Cys Asn Ala Ser Val Thr
Asn Ser 50 55 60 Val Lys Gly Thr Asn Ser Gly Glu Asn Leu Tyr Phe
Gln Gly Asp Pro65 70 75 80 Lys Ser Cys Asp Lys Thr His Thr Cys Pro
Pro Cys Pro Ala Pro Glu 85 90 95 Leu Leu Gly Gly Pro Ser Val Phe
Leu Phe Pro Pro Lys Pro Lys Asp 100 105 110 Thr Leu Met Ile Ser Arg
Thr Pro Glu Val Thr Cys Val Val Val Asp 115 120 125 Val Ser His Glu
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 130 135 140 Val Glu
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn145 150 155
160 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
165 170 175 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
Leu Pro 180 185 190 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln Pro Arg Glu 195 200 205 Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
Asp Glu Leu Thr Lys Asn 210 215 220 Gln Val Ser Leu Thr Cys Leu Val
Lys Gly Phe Tyr Pro Ser Asp Ile225 230 235 240 Ala Val Glu Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 245 250 255 Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 260 265 270 Leu
Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 275 280
285 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
290 295 300 Ser Leu Ser Pro Gly Lys305 310 38930DNAhomo sapiens
38atgccgctgc tgctactgct gcccctgctg tgggcagggg cgctagctat gctgcagatg
60gccggccagt gcagccagaa cgagtacttc gacagcctgc tgcacgcctg catcccctgc
120cagctgagat gcagcagcaa cacacctcct ctgacctgcc agagatactg
caacgccagc 180gtgaccaaca gcgtgaaggg caccaactcc ggagagaacc
tgtacttcca aggggatccc 240aaatcttgtg acaaaactca cacatgccca
ccgtgcccag cacctgaact cctgggggga 300ccgtcagtct tcctcttccc
cccaaaaccc aaggacaccc tcatgatctc ccggacccct 360gaggtcacat
gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg
420tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga
gcagtacaac 480agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc
aggactggct gaatggcaag 540gagtacaagt gcaaggtctc caacaaagcc
ctcccagccc ccatcgagaa aaccatctcc 600aaagccaaag ggcagccccg
agagccacag gtgtacaccc tgcccccatc ccgggatgag 660ctgaccaaga
accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc
720gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac
gcctcccgtg 780ctggactccg acggctcctt cttcctctac agcaagctca
ccgtggacaa gagcaggtgg 840cagcagggga acgtcttctc atgctccgtg
atgcatgagg ctctgcacaa ccactacacg 900cagaagagcc tctccctgtc
tccgggtaaa 93039307PRThomo sapiens 39Met Pro Leu Leu Leu Leu Leu
Pro Leu Leu Trp Ala Gly Ala Leu Ala1 5 10 15 Met Ala Gly Gln Cys
Ser Gln Asn Glu Tyr Phe Asp Ser Leu Leu His 20 25 30 Ala Cys Ile
Pro Cys Gln Leu Arg Cys Ser Ser Asn Thr Pro Pro Leu 35 40 45 Thr
Cys Gln Arg Tyr Cys Asn Ala Ser Val Thr Asn Ser Val Lys Gly 50 55
60 Thr Asn Ser Gly Glu Asn Leu Tyr Phe Gln Gly Asp Pro Lys Ser
Cys65 70 75 80 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu
Leu Leu Gly 85 90 95 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met 100 105 110 Ile Ser Arg Thr Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His 115 120 125 Glu Asp Pro Glu Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val 130 135 140 His Asn Ala Lys Thr
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr145 150 155 160 Arg Val
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 165 170 175
Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 180
185 190 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
Val 195 200 205 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn
Gln Val Ser 210 215 220 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
Asp Ile Ala Val Glu225 230 235 240 Trp Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro 245 250 255 Val Leu Asp Ser Asp Gly
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 260 265 270 Asp Lys Ser Arg
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 275 280 285 His Glu
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 290 295 300
Pro Gly Lys305 40921DNAhomo sapiens 40atgccgctgc tgctactgct
gcccctgctg tgggcagggg cgctagctat ggccggccag 60tgcagccaga acgagtactt
cgacagcctg ctgcacgcct gcatcccctg ccagctgaga 120tgcagcagca
acacacctcc tctgacctgc cagagatact gcaacgccag cgtgaccaac
180agcgtgaagg gcaccaactc cggagagaac ctgtacttcc aaggggatcc
caaatcttgt 240gacaaaactc acacatgccc accgtgccca gcacctgaac
tcctgggggg accgtcagtc 300ttcctcttcc ccccaaaacc caaggacacc
ctcatgatct cccggacccc tgaggtcaca 360tgcgtggtgg tggacgtgag
ccacgaagac cctgaggtca agttcaactg gtacgtggac 420ggcgtggagg
tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac
480cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa
ggagtacaag 540tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga
aaaccatctc caaagccaaa 600gggcagcccc gagagccaca ggtgtacacc
ctgcccccat cccgggatga gctgaccaag 660aaccaggtca gcctgacctg
cctggtcaaa ggcttctatc ccagcgacat cgccgtggag 720tgggagagca
atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc
780gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg
gcagcagggg 840aacgtcttct catgctccgt gatgcatgag gctctgcaca
accactacac gcagaagagc 900ctctccctgt ctccgggtaa a
92141306PRTcynomolgous macaque 41Met Pro Leu Leu Leu Leu Leu Pro
Leu Leu Trp Ala Gly Ala Leu Ala1 5 10 15 Met Ala Arg Gln Cys Ser
Gln Asn Glu Tyr Phe Asp Ser Leu Leu His 20 25 30 Asp Cys Lys Pro
Cys Gln Leu Arg Cys Ser Ser Thr Pro Pro Leu Thr 35 40 45 Cys Gln
Arg Tyr Cys Asn Ala Ser Met Thr Asn Ser Val Lys Gly Met 50 55 60
Asn Ser Gly Glu Asn Leu Tyr Phe Gln Gly Asp Pro Lys Ser Cys Asp65
70 75 80 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu
Gly Gly 85 90 95 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile 100 105 110 Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser His Glu 115 120 125 Asp Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His 130 135 140 Asn Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg145 150 155 160 Val Val Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 165 170 175 Glu
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 180 185
190 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
195 200 205 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val
Ser Leu 210 215 220 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
Ala Val Glu Trp225 230 235 240 Glu Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val 245 250 255 Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys Leu Thr Val Asp 260 265 270 Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys Ser Val Met His 275 280 285 Glu Ala Leu
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 290 295 300 Gly
Lys305 42918DNAcynomolgous macaque 42atgccgctgc tgctactgct
gcccctgctg tgggcagggg cgctagctat ggccagacag 60tgcagccaga acgagtactt
cgacagcctg ctgcacgact gcaagccctg ccagctgaga 120tgcagcagca
cacctcctct gacctgccag agatactgca acgccagcat gaccaacagc
180gtgaagggca tgaactccgg agagaacctg tacttccaag gggatcccaa
atcttgtgac 240aaaactcaca catgcccacc gtgcccagca cctgaactcc
tggggggacc gtcagtcttc 300ctcttccccc caaaacccaa ggacaccctc
atgatctccc ggacccctga ggtcacatgc 360gtggtggtgg acgtgagcca
cgaagaccct gaggtcaagt tcaactggta cgtggacggc 420gtggaggtgc
ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt
480gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga
gtacaagtgc 540aaggtctcca acaaagccct cccagccccc atcgagaaaa
ccatctccaa agccaaaggg 600cagccccgag agccacaggt gtacaccctg
cccccatccc gggatgagct gaccaagaac 660caggtcagcc tgacctgcct
ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 720gagagcaatg
ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac
780ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca
gcaggggaac 840gtcttctcat gctccgtgat gcatgaggct ctgcacaacc
actacacgca gaagagcctc 900tccctgtctc cgggtaaa 91843451PRTArtificial
SequenceHumansed antibody sequence 43Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Gly Thr Phe Ser Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155 160 Ser Trp
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180
185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
Pro Glu Leu Leu Gly225
230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg
Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu
Thr Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320 Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345
350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys
450 441353DNAArtificial SequenceHumansed antibody sequence
44caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg ccagcggcgg caccttcagc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
agagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcgc caggggcgcc 300atctacaacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agcgccagca
ccaagggccc cagcgtgttc cccctggccc ccagcagcaa gagcaccagc
420ggcggcacag ccgccctggg ctgcctggtg aaggactact tccccgaacc
ggtgaccgtg 480tcctggaaca gcggagccct gaccagcggc gtgcacacct
tccccgccgt gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg
accgtgccca gcagcagcct gggcacccag 600acctacatct gtaacgtgaa
ccacaagccc agcaacacca aggtggacaa gaaggtggag 660cccaagagct
gtgacaagac ccacacctgc cccccctgcc ctgcccccga gctgctggga
720ggccccagcg tgttcctgtt cccccccaag cctaaggaca ccctgatgat
cagcagaacc 780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg
accctgaggt gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat
gccaagacca agcccaggga ggagcagtac 900aacagcacct accgggtggt
gtccgtgctg accgtgctgc accaggattg gctgaacggc 960aaggagtaca
agtgtaaggt gtccaacaag gccctgcctg cccctatcga gaaaaccatc
1020agcaaggcca agggccagcc cagagagccc caggtgtaca ccctgccccc
tagcagagat 1080gagctgacca agaaccaggt gtccctgacc tgcctggtga
agggcttcta ccccagcgac 1140atcgccgtgg agtgggagag caacggccag
cccgagaaca actacaagac caccccccct 1200gtgctggaca gcgatggcag
cttcttcctg tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg
gcaacgtgtt cagctgctcc gtgatgcacg aggccctgca caatcactac
1320acccagaaga gcctgagcct gtcccctggc aag 135345451PRTArtificial
SequenceHumansed antibody sequence 45Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155 160 Ser Trp
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180
185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
Pro Glu Leu Leu Gly225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly305
310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400 Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425
430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445 Pro Gly Lys 450 461353DNAArtificial SequenceHumansed
antibody sequence 46caggtgcagc tggtccagag cggcgccgaa gtgaagaagc
ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta caccttcacc aactactgga
tgcactgggt gaggcaggcc 120cccggacagg gcctggagtg gatgggcgcc
acctacaggg gccacagcga cacctactac 180aaccagaagt tcaagggccg
ggtgaccatc accgccgaca agagcaccag caccgcctac 240atggaactga
gcagcctcag gagcgaggac accgctgtgt attactgcgc caggggcgcc
300atctacaacg gctacgacgt gctggacaac tggggccagg gcacactagt
gaccgtgtcc 360agcgccagca ccaagggccc cagcgtgttc cccctggccc
ccagcagcaa gagcaccagc 420ggcggcacag ccgccctggg ctgcctggtg
aaggactact tccccgaacc ggtgaccgtg 480tcctggaaca gcggagccct
gaccagcggc gtgcacacct tccccgccgt gctgcagagc 540agcggcctgt
acagcctgag cagcgtggtg accgtgccca gcagcagcct gggcacccag
600acctacatct gtaacgtgaa ccacaagccc agcaacacca aggtggacaa
gaaggtggag 660cccaagagct gtgacaagac ccacacctgc cccccctgcc
ctgcccccga gctgctggga 720ggccccagcg tgttcctgtt cccccccaag
cctaaggaca ccctgatgat cagcagaacc 780cccgaggtga cctgtgtggt
ggtggatgtg agccacgagg accctgaggt gaagttcaac 840tggtacgtgg
acggcgtgga ggtgcacaat gccaagacca agcccaggga ggagcagtac
900aacagcacct accgggtggt gtccgtgctg accgtgctgc accaggattg
gctgaacggc 960aaggagtaca agtgtaaggt gtccaacaag gccctgcctg
cccctatcga gaaaaccatc 1020agcaaggcca agggccagcc cagagagccc
caggtgtaca ccctgccccc tagcagagat 1080gagctgacca agaaccaggt
gtccctgacc tgcctggtga agggcttcta ccccagcgac 1140atcgccgtgg
agtgggagag caacggccag cccgagaaca actacaagac caccccccct
1200gtgctggaca gcgatggcag cttcttcctg tacagcaagc tgaccgtgga
caagagcaga 1260tggcagcagg gcaacgtgtt cagctgctcc gtgatgcacg
aggccctgca caatcactac 1320acccagaaga gcctgagcct gtcccctggc aag
135347451PRTArtificial SequenceHumansed antibody sequence 47Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20
25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asn
Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150
155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235 240 Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275
280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395
400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 481353DNAArtificial
SequenceHumansed antibody sequence 48caggtgcagc tggtccagag
cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta
caccttcacc aactactgga tgcactgggt gaggcaggcc 120cccggacagg
gcctggagtg gatgggcgcc acctacaggg gccacagcga cacctactac
180aaccagaagt tcaagggccg ggtgaccatc accgccgaca agagcaccag
caccgcctac 240atggaactga gcagcctcag gagcgaggac accgctgtgt
attactgcac caggggcgcc 300atctacaacg gctacgacgt gctggacaac
tggggccagg gcacactagt gaccgtgtcc 360agcgccagca ccaagggccc
cagcgtgttc cccctggccc ccagcagcaa gagcaccagc 420ggcggcacag
ccgccctggg ctgcctggtg aaggactact tccccgaacc ggtgaccgtg
480tcctggaaca gcggagccct gaccagcggc gtgcacacct tccccgccgt
gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg accgtgccca
gcagcagcct gggcacccag 600acctacatct gtaacgtgaa ccacaagccc
agcaacacca aggtggacaa gaaggtggag 660cccaagagct gtgacaagac
ccacacctgc cccccctgcc ctgcccccga gctgctggga 720ggccccagcg
tgttcctgtt cccccccaag cctaaggaca ccctgatgat cagcagaacc
780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg accctgaggt
gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat gccaagacca
agcccaggga ggagcagtac 900aacagcacct accgggtggt gtccgtgctg
accgtgctgc accaggattg gctgaacggc 960aaggagtaca agtgtaaggt
gtccaacaag gccctgcctg cccctatcga gaaaaccatc 1020agcaaggcca
agggccagcc cagagagccc caggtgtaca ccctgccccc tagcagagat
1080gagctgacca agaaccaggt gtccctgacc tgcctggtga agggcttcta
ccccagcgac 1140atcgccgtgg agtgggagag caacggccag cccgagaaca
actacaagac caccccccct 1200gtgctggaca gcgatggcag cttcttcctg
tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg gcaacgtgtt
cagctgctcc gtgatgcacg aggccctgca caatcactac 1320acccagaaga
gcctgagcct gtcccctggc aag 135349451PRTArtificial SequenceHumansed
antibody sequence 49Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Gly Ser Gly
Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala
Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Thr Tyr Arg Gly
His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val
Thr Ile Thr Ala Asp Thr Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu
Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Thr Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val Leu Asp Asn Trp Gly 100
105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly
Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro Val Thr Val145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225
230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg
Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu
Thr Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320 Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345
350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys
450 501353DNAArtificial SequenceHumansed antibody sequence
50caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg gcagcggcta caccttcacc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
cgagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcac caggggcgcc 300atctacaacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agcgccagca
ccaagggccc cagcgtgttc cccctggccc ccagcagcaa gagcaccagc
420ggcggcacag ccgccctggg ctgcctggtg aaggactact tccccgaacc
ggtgaccgtg 480tcctggaaca gcggagccct gaccagcggc gtgcacacct
tccccgccgt gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg
accgtgccca gcagcagcct gggcacccag 600acctacatct gtaacgtgaa
ccacaagccc agcaacacca aggtggacaa gaaggtggag 660cccaagagct
gtgacaagac ccacacctgc cccccctgcc ctgcccccga gctgctggga
720ggccccagcg tgttcctgtt cccccccaag cctaaggaca ccctgatgat
cagcagaacc 780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg
accctgaggt gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat
gccaagacca agcccaggga ggagcagtac 900aacagcacct accgggtggt
gtccgtgctg accgtgctgc accaggattg gctgaacggc 960aaggagtaca
agtgtaaggt gtccaacaag gccctgcctg cccctatcga gaaaaccatc
1020agcaaggcca agggccagcc cagagagccc caggtgtaca ccctgccccc
tagcagagat 1080gagctgacca agaaccaggt gtccctgacc tgcctggtga
agggcttcta ccccagcgac 1140atcgccgtgg agtgggagag caacggccag
cccgagaaca actacaagac caccccccct 1200gtgctggaca gcgatggcag
cttcttcctg tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg
gcaacgtgtt cagctgctcc gtgatgcacg aggccctgca caatcactac
1320acccagaaga gcctgagcct gtcccctggc aag 135351451PRTArtificial
SequenceHumansed antibody sequence 51Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Thr Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asn Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155 160 Ser Trp
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180
185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
Pro Glu Leu Leu Gly225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly305
310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400 Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425
430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445 Pro Gly Lys 450 521353DNAArtificial SequenceHumansed
antibody sequence 52caggtgcagc tggtccagag cggcgccgaa gtgaagaagc
ccggcagctc cgtgaaagtg 60agctgcaagg gcagcggcta caccttcacc aactactgga
tgcactgggt gaggcaggcc 120cccggacagg gcctggagtg gatcggcgcc
acctacaggg gccacagcga cacctactac 180aaccagaagt tcaagggccg
ggcgaccctc accgccgaca cgagcaccag caccgcctac 240atggaactga
gcagcctcag gagcgaggac accgctgtgt attactgcac caggggcgcc
300atctacaacg gctacgacgt gctggacaac tggggccagg gcacactagt
gaccgtgtcc 360agcgccagca ccaagggccc cagcgtgttc cccctggccc
ccagcagcaa gagcaccagc 420ggcggcacag ccgccctggg ctgcctggtg
aaggactact tccccgaacc ggtgaccgtg 480tcctggaaca gcggagccct
gaccagcggc gtgcacacct tccccgccgt gctgcagagc 540agcggcctgt
acagcctgag cagcgtggtg accgtgccca gcagcagcct gggcacccag
600acctacatct gtaacgtgaa ccacaagccc agcaacacca aggtggacaa
gaaggtggag 660cccaagagct gtgacaagac ccacacctgc cccccctgcc
ctgcccccga gctgctggga 720ggccccagcg tgttcctgtt cccccccaag
cctaaggaca ccctgatgat cagcagaacc 780cccgaggtga cctgtgtggt
ggtggatgtg agccacgagg accctgaggt gaagttcaac 840tggtacgtgg
acggcgtgga ggtgcacaat gccaagacca agcccaggga ggagcagtac
900aacagcacct accgggtggt gtccgtgctg accgtgctgc accaggattg
gctgaacggc 960aaggagtaca agtgtaaggt gtccaacaag gccctgcctg
cccctatcga gaaaaccatc 1020agcaaggcca agggccagcc cagagagccc
caggtgtaca ccctgccccc tagcagagat 1080gagctgacca agaaccaggt
gtccctgacc tgcctggtga agggcttcta ccccagcgac 1140atcgccgtgg
agtgggagag caacggccag cccgagaaca actacaagac caccccccct
1200gtgctggaca gcgatggcag cttcttcctg tacagcaagc tgaccgtgga
caagagcaga 1260tggcagcagg gcaacgtgtt cagctgctcc gtgatgcacg
aggccctgca caatcactac 1320acccagaaga gcctgagcct gtcccctggc aag
135353451PRTArtificial SequenceHumansed antibody sequence 53Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asn Tyr 20
25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Thr Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asp
Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150
155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235 240 Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275
280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395
400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 541353DNAArtificial
SequenceHumansed antibody sequence 54caggtgcagc tggtccagag
cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg gcagcggcta
caccttcacc aactactgga tgcactgggt gaggcaggcc 120cccggacagg
gcctggagtg gatgggcgcc acctacaggg gccacagcga cacctactac
180aaccagaagt tcaagggccg ggtgaccatc accgccgaca cgagcaccag
caccgcctac 240atggaactga gcagcctcag gagcgaggac accgctgtgt
attactgcac caggggcgcc 300atctacgacg gctacgacgt gctggacaac
tggggccagg gcacactagt gaccgtgtcc 360agcgccagca ccaagggccc
cagcgtgttc cccctggccc ccagcagcaa gagcaccagc 420ggcggcacag
ccgccctggg ctgcctggtg aaggactact tccccgaacc ggtgaccgtg
480tcctggaaca gcggagccct gaccagcggc gtgcacacct tccccgccgt
gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg accgtgccca
gcagcagcct gggcacccag 600acctacatct gtaacgtgaa ccacaagccc
agcaacacca aggtggacaa gaaggtggag 660cccaagagct gtgacaagac
ccacacctgc cccccctgcc ctgcccccga gctgctggga 720ggccccagcg
tgttcctgtt cccccccaag cctaaggaca ccctgatgat cagcagaacc
780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg accctgaggt
gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat gccaagacca
agcccaggga ggagcagtac 900aacagcacct accgggtggt gtccgtgctg
accgtgctgc accaggattg gctgaacggc 960aaggagtaca agtgtaaggt
gtccaacaag gccctgcctg cccctatcga gaaaaccatc 1020agcaaggcca
agggccagcc cagagagccc caggtgtaca ccctgccccc tagcagagat
1080gagctgacca agaaccaggt gtccctgacc tgcctggtga agggcttcta
ccccagcgac 1140atcgccgtgg agtgggagag caacggccag cccgagaaca
actacaagac caccccccct 1200gtgctggaca gcgatggcag cttcttcctg
tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg gcaacgtgtt
cagctgctcc gtgatgcacg aggccctgca caatcactac 1320acccagaaga
gcctgagcct gtcccctggc aag 135355451PRTArtificial SequenceHumansed
antibody sequence 55Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Gly Thr Phe Ser Asn Tyr 20 25 30 Trp Met His Trp Val Arg Gln Ala
Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Ala Thr Tyr Arg Gly
His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val
Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu
Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Ala Arg Gly Ala Ile Tyr Asp Gly Tyr Asp Val Leu Asp Asn Trp Gly 100
105 110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro
Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly
Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro Val Thr Val145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser
Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro
Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225
230 235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
Val Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg
Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu
Thr Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320 Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345
350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
355 360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys
450 561353DNAArtificial SequenceHumansed antibody sequence
56caggtgcagc tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg
60agctgcaagg ccagcggcgg caccttcagc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
agagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcgc caggggcgcc 300atctacgacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agcgccagca
ccaagggccc cagcgtgttc cccctggccc ccagcagcaa gagcaccagc
420ggcggcacag ccgccctggg ctgcctggtg aaggactact tccccgaacc
ggtgaccgtg 480tcctggaaca gcggagccct gaccagcggc gtgcacacct
tccccgccgt gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg
accgtgccca gcagcagcct gggcacccag 600acctacatct gtaacgtgaa
ccacaagccc agcaacacca aggtggacaa gaaggtggag 660cccaagagct
gtgacaagac ccacacctgc cccccctgcc ctgcccccga gctgctggga
720ggccccagcg tgttcctgtt cccccccaag cctaaggaca ccctgatgat
cagcagaacc 780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg
accctgaggt gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat
gccaagacca agcccaggga ggagcagtac 900aacagcacct accgggtggt
gtccgtgctg accgtgctgc accaggattg gctgaacggc 960aaggagtaca
agtgtaaggt gtccaacaag gccctgcctg cccctatcga gaaaaccatc
1020agcaaggcca agggccagcc cagagagccc caggtgtaca ccctgccccc
tagcagagat 1080gagctgacca agaaccaggt gtccctgacc tgcctggtga
agggcttcta ccccagcgac 1140atcgccgtgg agtgggagag caacggccag
cccgagaaca actacaagac caccccccct 1200gtgctggaca gcgatggcag
cttcttcctg tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg
gcaacgtgtt cagctgctcc gtgatgcacg aggccctgca caatcactac
1320acccagaaga gcctgagcct gtcccctggc aag 135357451PRTArtificial
SequenceHumansed antibody sequence 57Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala
Arg Gly Ala Ile Tyr Asp Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105
110 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
115 120 125 Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
Thr Ala 130 135 140 Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val145 150 155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val 180 185 190 Pro Ser Ser Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His 195 200 205 Lys Pro Ser
Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230
235 240 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met 245 250 255 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His 260 265 270 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val 275 280 285 His Asn Ala Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr 290 295 300 Arg Val Val Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320 Lys Glu Tyr Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350
Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355
360 365 Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu 370 375 380 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro385 390 395 400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val 405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met 420 425 430 His Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450
581353DNAArtificial SequenceHumansed antibody sequence 58caggtgcagc
tggtccagag cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg
ccagcggcta caccttcacc aactactgga tgcactgggt gaggcaggcc
120cccggacagg gcctggagtg gatgggcgcc acctacaggg gccacagcga
cacctactac 180aaccagaagt tcaagggccg ggtgaccatc accgccgaca
agagcaccag caccgcctac 240atggaactga gcagcctcag gagcgaggac
accgctgtgt attactgcgc caggggcgcc 300atctacgacg gctacgacgt
gctggacaac tggggccagg gcacactagt gaccgtgtcc 360agcgccagca
ccaagggccc cagcgtgttc cccctggccc ccagcagcaa gagcaccagc
420ggcggcacag ccgccctggg ctgcctggtg aaggactact tccccgaacc
ggtgaccgtg 480tcctggaaca gcggagccct gaccagcggc gtgcacacct
tccccgccgt gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg
accgtgccca gcagcagcct gggcacccag 600acctacatct gtaacgtgaa
ccacaagccc agcaacacca aggtggacaa gaaggtggag 660cccaagagct
gtgacaagac ccacacctgc cccccctgcc ctgcccccga gctgctggga
720ggccccagcg tgttcctgtt cccccccaag cctaaggaca ccctgatgat
cagcagaacc 780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg
accctgaggt gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat
gccaagacca agcccaggga ggagcagtac 900aacagcacct accgggtggt
gtccgtgctg accgtgctgc accaggattg gctgaacggc 960aaggagtaca
agtgtaaggt gtccaacaag gccctgcctg cccctatcga gaaaaccatc
1020agcaaggcca agggccagcc cagagagccc caggtgtaca ccctgccccc
tagcagagat 1080gagctgacca agaaccaggt gtccctgacc tgcctggtga
agggcttcta ccccagcgac 1140atcgccgtgg agtgggagag caacggccag
cccgagaaca actacaagac caccccccct 1200gtgctggaca gcgatggcag
cttcttcctg tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg
gcaacgtgtt cagctgctcc gtgatgcacg aggccctgca caatcactac
1320acccagaaga gcctgagcct gtcccctggc aag 135359451PRTArtificial
SequenceHumansed antibody sequence 59Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asp Gly Tyr Asp Val
Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro Leu Ala Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala Leu Gly Cys Leu
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150 155 160 Ser Trp
Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 165 170 175
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 180
185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro
Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala
Pro Glu Leu Leu Gly225 230 235 240 Gly Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His 260 265 270 Glu Asp Pro Glu
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285 His Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly305
310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu
Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400 Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415 Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425
430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445 Pro Gly Lys 450 601353DNAArtificial SequenceHumansed
antibody sequence 60caggtgcagc tggtccagag cggcgccgaa gtgaagaagc
ccggcagctc cgtgaaagtg 60agctgcaagg ccagcggcta caccttcacc aactactgga
tgcactgggt gaggcaggcc 120cccggacagg gcctggagtg gatgggcgcc
acctacaggg gccacagcga cacctactac 180aaccagaagt tcaagggccg
ggtgaccatc accgccgaca agagcaccag caccgcctac 240atggaactga
gcagcctcag gagcgaggac accgctgtgt attactgcac caggggcgcc
300atctacgacg gctacgacgt gctggacaac tggggccagg gcacactagt
gaccgtgtcc 360agcgccagca ccaagggccc cagcgtgttc cccctggccc
ccagcagcaa gagcaccagc 420ggcggcacag ccgccctggg ctgcctggtg
aaggactact tccccgaacc ggtgaccgtg 480tcctggaaca gcggagccct
gaccagcggc gtgcacacct tccccgccgt gctgcagagc 540agcggcctgt
acagcctgag cagcgtggtg accgtgccca gcagcagcct gggcacccag
600acctacatct gtaacgtgaa ccacaagccc agcaacacca aggtggacaa
gaaggtggag 660cccaagagct gtgacaagac ccacacctgc cccccctgcc
ctgcccccga gctgctggga 720ggccccagcg tgttcctgtt cccccccaag
cctaaggaca ccctgatgat cagcagaacc 780cccgaggtga cctgtgtggt
ggtggatgtg agccacgagg accctgaggt gaagttcaac 840tggtacgtgg
acggcgtgga ggtgcacaat gccaagacca agcccaggga ggagcagtac
900aacagcacct accgggtggt gtccgtgctg accgtgctgc accaggattg
gctgaacggc 960aaggagtaca agtgtaaggt gtccaacaag gccctgcctg
cccctatcga gaaaaccatc 1020agcaaggcca agggccagcc cagagagccc
caggtgtaca ccctgccccc tagcagagat 1080gagctgacca agaaccaggt
gtccctgacc tgcctggtga agggcttcta ccccagcgac 1140atcgccgtgg
agtgggagag caacggccag cccgagaaca actacaagac caccccccct
1200gtgctggaca gcgatggcag cttcttcctg tacagcaagc tgaccgtgga
caagagcaga 1260tggcagcagg gcaacgtgtt cagctgctcc gtgatgcacg
aggccctgca caatcactac 1320acccagaaga gcctgagcct gtcccctggc aag
135361451PRTArtificial SequenceHumansed antibody sequence 61Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe Thr Asn Tyr 20
25 30 Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Ile 35 40 45 Gly Ala Thr Tyr Arg Gly His Ser Asp Thr Tyr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Ala Asp Thr Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Thr Arg Gly Ala Ile Tyr Asp
Gly Tyr Asp Val Leu Asp Asn Trp Gly 100 105 110 Gln Gly Thr Leu Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 115 120 125 Val Phe Pro
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala 130 135 140 Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val145 150
155 160 Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala 165 170 175 Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val 180 185 190 Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His 195 200 205 Lys Pro Ser Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys 210 215 220 Asp Lys Thr His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235 240 Gly Pro Ser Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255 Ile Ser
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270
Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275
280 285 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr 290 295 300 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly305 310 315 320 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile 325 330 335 Glu Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val 340 345 350 Tyr Thr Leu Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 355 360 365 Leu Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380 Trp Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395
400 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
Val Met 420 425 430 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser 435 440 445 Pro Gly Lys 450 621353DNAArtificial
SequenceHumansed antibody sequence 62caggtgcagc tggtccagag
cggcgccgaa gtgaagaagc ccggcagctc cgtgaaagtg 60agctgcaagg gcagcggcta
caccttcacc aactactgga tgcactgggt gaggcaggcc 120cccggacagg
gcctggagtg gatcggcgcc acctacaggg gccacagcga cacctactac
180aaccagaagt tcaagggccg ggcgaccctc accgccgaca cgagcaccag
caccgcctac 240atggaactga gcagcctcag gagcgaggac accgctgtgt
attactgcac caggggcgcc 300atctacgacg gctacgacgt gctggacaac
tggggccagg gcacactagt gaccgtgtcc 360agcgccagca ccaagggccc
cagcgtgttc cccctggccc ccagcagcaa gagcaccagc 420ggcggcacag
ccgccctggg ctgcctggtg aaggactact tccccgaacc ggtgaccgtg
480tcctggaaca gcggagccct gaccagcggc gtgcacacct tccccgccgt
gctgcagagc 540agcggcctgt acagcctgag cagcgtggtg accgtgccca
gcagcagcct gggcacccag 600acctacatct gtaacgtgaa ccacaagccc
agcaacacca aggtggacaa gaaggtggag 660cccaagagct gtgacaagac
ccacacctgc cccccctgcc ctgcccccga gctgctggga 720ggccccagcg
tgttcctgtt cccccccaag cctaaggaca ccctgatgat cagcagaacc
780cccgaggtga cctgtgtggt ggtggatgtg agccacgagg accctgaggt
gaagttcaac 840tggtacgtgg acggcgtgga ggtgcacaat gccaagacca
agcccaggga ggagcagtac 900aacagcacct accgggtggt gtccgtgctg
accgtgctgc accaggattg gctgaacggc 960aaggagtaca agtgtaaggt
gtccaacaag gccctgcctg cccctatcga gaaaaccatc 1020agcaaggcca
agggccagcc cagagagccc caggtgtaca ccctgccccc tagcagagat
1080gagctgacca agaaccaggt gtccctgacc tgcctggtga agggcttcta
ccccagcgac 1140atcgccgtgg agtgggagag caacggccag cccgagaaca
actacaagac caccccccct 1200gtgctggaca gcgatggcag cttcttcctg
tacagcaagc tgaccgtgga caagagcaga 1260tggcagcagg gcaacgtgtt
cagctgctcc gtgatgcacg aggccctgca caatcactac 1320acccagaaga
gcctgagcct gtcccctggc aag 135363214PRTArtificial SequenceHumansed
antibody sequence 63Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser
Gln Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Asn Leu
His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80 Glu Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Arg Lys Leu Pro Trp 85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100
105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
Ser Gly Asn Ser Gln145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser
Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val
Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn
Arg Gly Glu Cys 210 64642DNAArtificial SequenceHumansed antibody
sequence 64gacatccaga tgacccagag ccctagctca ctgagcgcca gcgtgggcga
cagggtgacc 60attacctgct ccgccagcca ggacatcagc aactacctga actggtacca
gcagaagccc 120ggcaaggccc ccaagctgct gatctactac acctccaacc
tgcactccgg cgtgcccagc 180aggttcagcg gaagcggcag cggcaccgat
ttcaccctga ccatctccag cctgcagccc 240gaggacttcg ccacctacta
ctgccagcag tacaggaagc tcccctggac tttcggccag 300ggcaccaaac
tggagatcaa gcgtacggtg gccgccccca gcgtgttcat cttccccccc
360agcgatgagc agctgaagag cggcaccgcc agcgtggtgt gtctgctgaa
caacttctac 420ccccgggagg ccaaggtgca gtggaaggtg gacaatgccc
tgcagagcgg caacagccag 480gagagcgtga ccgagcagga cagcaaggac
tccacctaca gcctgagcag caccctgacc 540ctgagcaagg ccgactacga
gaagcacaag gtgtacgcct gtgaggtgac ccaccagggc 600ctgtccagcc
ccgtgaccaa gagcttcaac cggggcgagt gc 64265214PRTArtificial
SequenceHumansed antibody sequence 65Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15 Asp Arg Val Thr Ile
Thr Cys Ser Ala Ser Gln Asp Ile Ser Asn Tyr 20 25
30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45 Tyr Tyr Thr Ser Asn Leu His Ser 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 Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
Tyr Arg Lys Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val
Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155
160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys
Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210
66642DNAArtificial SequenceHumansed antibody sequence 66gacatccaga
tgacccagag ccctagctca ctgagcgcca gcgtgggcga cagggtgacc 60attacctgct
ccgccagcca ggacatcagc aactacctga actggtacca gcagaagccc
120ggcaaggccc ccaagctgct gatctactac acctccaacc tgcactccgg
cgtgcccagc 180aggttcagcg gaagcggcag cggcaccgat tacaccctga
ccatctccag cctgcagccc 240gaggacttcg ccacctacta ctgccagcag
tacaggaagc tcccctggac tttcggccag 300ggcaccaaac tggagatcaa
gcgtacggtg gccgccccca gcgtgttcat cttccccccc 360agcgatgagc
agctgaagag cggcaccgcc agcgtggtgt gtctgctgaa caacttctac
420ccccgggagg ccaaggtgca gtggaaggtg gacaatgccc tgcagagcgg
caacagccag 480gagagcgtga ccgagcagga cagcaaggac tccacctaca
gcctgagcag caccctgacc 540ctgagcaagg ccgactacga gaagcacaag
gtgtacgcct gtgaggtgac ccaccagggc 600ctgtccagcc ccgtgaccaa
gagcttcaac cggggcgagt gc 64267214PRTArtificial SequenceHumansed
antibody sequence 67Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser
Gln Asp Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Glu Leu Val Ile 35 40 45 Tyr Tyr Thr Ser Asn Leu
His Ser 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 Pro65 70 75 80 Glu Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Arg Lys Leu Pro Trp 85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100
105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser
Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
Ser Gly Asn Ser Gln145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser
Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser
Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val
Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn
Arg Gly Glu Cys 210 68642DNAArtificial SequenceHumansed antibody
sequence 68gacatccagc tgacccagag ccctagctca ctgagcgcca gcgtgggcga
cagggtgacc 60attacctgct ccgccagcca ggacatcagc aactacctga actggtacca
gcagaagccc 120ggcaaggccc ccgagctggt gatctactac acctccaacc
tgcactccgg cgtgcccagc 180aggttcagcg gaagcggcag cggcaccgat
tacaccctga ccatctccag cctgcagccc 240gaggacttcg ccacctacta
ctgccagcag tacaggaagc tcccctggac tttcggccag 300ggcaccaaac
tggagatcaa gcgtacggtg gccgccccca gcgtgttcat cttccccccc
360agcgatgagc agctgaagag cggcaccgcc agcgtggtgt gtctgctgaa
caacttctac 420ccccgggagg ccaaggtgca gtggaaggtg gacaatgccc
tgcagagcgg caacagccag 480gagagcgtga ccgagcagga cagcaaggac
tccacctaca gcctgagcag caccctgacc 540ctgagcaagg ccgactacga
gaagcacaag gtgtacgcct gtgaggtgac ccaccagggc 600ctgtccagcc
ccgtgaccaa gagcttcaac cggggcgagt gc 64269115PRTmus musculus 69Glu
Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala1 5 10
15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
20 25 30 Tyr Met Lys Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu
Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr
Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys
Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser
Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Gly Tyr Glu Phe
Val Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ala 115
70345DNAmus musculus 70gaggtccagt tgcaacaatc tggacctgag ctggtgaagc
ctggggcttc agtgaagata 60tcctgtaagg cttctggata cacattcact gactactaca
tgaagtgggt gaagcagagc 120catggaaaga gccttgagtg gattggagag
atttatccta ataatggtgg tattacctac 180aaccagaagt tcaagggcaa
ggccacattg actgtagaca agtcctccag cacagcctac 240atggagctcc
gcagcctgac atctgaggac tctgcagtct attactgtgc aaatggttac
300gagtttgttt actggggcca agggactctg gtcactgtct ctgca 34571108PRTmus
musculus 71Asp Ile Gln Met Thr Gln Thr Ala Ser Ser Leu Ser Ala Ser
Leu Gly1 5 10 15 Asp Arg Val Thr Ile Ser Cys Ser Ala Ser Gln Gly
Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly
Thr Val Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser
Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp
Tyr Ser Leu Thr Ile Ser Asn Leu Glu Pro65 70 75 80 Glu Asp Ile Ala
Thr Tyr Tyr Cys Gln Gln Tyr Ser Lys Leu Pro Trp 85 90 95 Thr Phe
Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 100 105 72324DNAmus
musculus 72gatatccaga tgacacagac tgcatcctcc ctgtctgcct ctctgggaga
cagagtcacc 60atcagttgca gtgcaagtca gggcattagc aattatttaa actggtatca
gcagaaacca 120gatggaactg ttaaactcct gatctattac acatcaagtt
tacactcagg agtcccatca 180aggttcagtg gcagtgggtc tgggacagat
tattctctca ccatcagcaa cctggaacct 240gaagatattg ccacttacta
ttgtcagcag tatagtaagc ttccgtggac gttcggtgga 300ggcaccaagc
tggaaatcaa acgg 32473445PRTArtificial SequenceChimeric antibody
sequence 73Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro
Gly Ala1 5 10 15 Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Asp Tyr 20 25 30 Tyr Met Lys Trp Val Lys Gln Ser His Gly
Lys Ser Leu Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Asn Asn Gly
Gly Ile Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu
Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Arg
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Asn
Gly Tyr Glu Phe Val Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110
Val Ser Ala Ala Lys Thr Thr Ala 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 Ala145 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 Asp Lys Thr His Thr Cys 210 215 220 Pro Pro
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu225 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 Lys305 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 Gly385 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 741335DNAArtificial
SequenceChimeric antibody sequence 74gaggtccagt tgcaacaatc
tggacctgag ctggtgaagc ctggggcttc agtgaagata 60tcctgtaagg cttctggata
cacattcact gactactaca tgaagtgggt gaagcagagc 120catggaaaga
gccttgagtg gattggagag atttatccta ataatggtgg tattacctac
180aaccagaagt tcaagggcaa ggccacattg actgtagaca agtcctccag
cacagcctac 240atggagctcc gcagcctgac atctgaggac tctgcagtct
attactgtgc aaatggttac 300gagtttgttt actggggcca agggactctg
gtcactgtct ctgcagccaa aacaacagcc 360cccagcgtgt tccccctggc
ccccagcagc aagagcacca gcggcggcac agccgccctg 420ggctgcctgg
tgaaggacta cttccccgaa ccggtgaccg tgtcctggaa cagcggagcc
480ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct
gtacagcctg 540agcagcgtgg tgaccgtgcc cagcagcagc ctgggcaccc
agacctacat ctgtaacgtg 600aaccacaagc ccagcaacac caaggtggac
aagaaggtgg agcccaagag ctgtgacaag 660acccacacct gccccccctg
ccctgccccc gagctgctgg gaggccccag cgtgttcctg 720ttccccccca
agcctaagga caccctgatg atcagcagaa cccccgaggt gacctgtgtg
780gtggtggatg tgagccacga ggaccctgag gtgaagttca actggtacgt
ggacggcgtg 840gaggtgcaca atgccaagac caagcccagg gaggagcagt
acaacagcac ctaccgggtg 900gtgtccgtgc tgaccgtgct gcaccaggat
tggctgaacg gcaaggagta caagtgtaag 960gtgtccaaca aggccctgcc
tgcccctatc gagaaaacca tcagcaaggc caagggccag 1020cccagagagc
cccaggtgta caccctgccc cctagcagag atgagctgac caagaaccag
1080gtgtccctga cctgcctggt gaagggcttc taccccagcg acatcgccgt
ggagtgggag 1140agcaacggcc agcccgagaa caactacaag accacccccc
ctgtgctgga cagcgatggc 1200agcttcttcc tgtacagcaa gctgaccgtg
gacaagagca gatggcagca gggcaacgtg 1260ttcagctgct ccgtgatgca
cgaggccctg cacaatcact acacccagaa gagcctgagc 1320ctgtcccctg gcaag
133575214PRTArtificial SequenceChimeric antibody sequence 75Asp Ile
Gln Met Thr Gln Thr Ala Ser Ser Leu Ser Ala Ser Leu Gly1 5 10 15
Asp Arg Val Thr Ile Ser Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr 20
25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu
Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser Gly Val Pro Ser Arg
Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile
Ser Asn Leu Glu Pro65 70 75 80 Glu Asp Ile Ala Thr Tyr Tyr Cys Gln
Gln Tyr Ser Lys Leu Pro Trp 85 90 95 Thr Phe Gly Gly Gly Thr Lys
Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile
Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser
Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys
Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150
155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210
76642DNAArtificial SequenceChimeric antibody sequence 76gatatccaga
tgacacagac tgcatcctcc ctgtctgcct ctctgggaga cagagtcacc 60atcagttgca
gtgcaagtca gggcattagc aattatttaa actggtatca gcagaaacca
120gatggaactg ttaaactcct gatctattac acatcaagtt tacactcagg
agtcccatca 180aggttcagtg gcagtgggtc tgggacagat tattctctca
ccatcagcaa cctggaacct 240gaagatattg ccacttacta ttgtcagcag
tatagtaagc ttccgtggac gttcggtgga 300ggcaccaagc tggagctgaa
acgtacggtg gccgccccca gcgtgttcat cttccccccc 360agcgatgagc
agctgaagag cggcaccgcc agcgtggtgt gtctgctgaa caacttctac
420ccccgggagg ccaaggtgca gtggaaggtg gacaatgccc tgcagagcgg
caacagccag 480gagagcgtga ccgagcagga cagcaaggac tccacctaca
gcctgagcag caccctgacc 540ctgagcaagg ccgactacga gaagcacaag
gtgtacgcct gtgaggtgac ccaccagggc 600ctgtccagcc ccgtgaccaa
gagcttcaac cggggcgagt gc 64277115PRTArtificial SequenceHumansed
antibody sequence 77Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Gly Thr Phe Ser Asp Tyr 20 25 30 Tyr Met Lys Trp Val Arg Gln Ala
Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Glu Ile Tyr Pro Asn
Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val
Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu
Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Ala Arg Gly Tyr Glu Phe Val Tyr Trp Gly Gln Gly Thr Leu Val Thr 100
105 110 Val Ser Ser 115 78345DNAArtificial SequenceHumansed
antibody sequence 78caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc
ccggctccag cgtgaaggtg 60agctgcaagg ctagcggcgg caccttcagc gactactaca
tgaagtgggt gaggcaggcc 120cccggccagg gactggagtg gatgggcgag
atctacccca acaacggggg catcacctac 180aaccagaagt tcaagggcag
ggtgaccatc accgccgaca aaagcaccag caccgcctac 240atggaactga
gcagcctgag gagcgaggac accgccgtgt actactgcgc caggggctac
300gagttcgtgt attggggcca gggcacacta gtgaccgtgt ccagc
34579115PRTArtificial SequenceHumansed antibody sequence 79Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20
25 30 Tyr Met Lys Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45 Gly Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Tyr Glu Phe Val
Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115
80345DNAArtificial SequenceHumansed antibody sequence 80caggtgcagc
tggtgcagag cggcgccgaa gtgaagaagc ccggctccag cgtgaaggtg 60agctgcaagg
ctagcggcta caccttcacc gactactaca tgaagtgggt gaggcaggcc
120cccggccagg gactggagtg gatgggcgag atctacccca acaacggggg
catcacctac 180aaccagaagt tcaagggcag ggtgaccatc accgccgaca
aaagcaccag caccgcctac 240atggaactga gcagcctgag gagcgaggac
accgccgtgt actactgcgc caggggctac 300gagttcgtgt attggggcca
gggcacacta gtgaccgtgt ccagc 34581115PRTArtificial SequenceHumansed
antibody sequence 81Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys Trp Val Arg Gln Ala
Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Glu Ile Tyr Pro Asn
Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val
Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu
Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Ala Asn Gly Tyr Glu Phe Val Tyr Trp Gly Gln Gly Thr Leu Val Thr 100
105 110 Val Ser Ser 115 82345DNAArtificial SequenceHumansed
antibody sequence 82caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc
ccggctccag cgtgaaggtg 60agctgcaagg ctagcggcta caccttcacc gactactaca
tgaagtgggt gaggcaggcc 120cccggccagg gactggagtg gatgggcgag
atctacccca acaacggggg catcacctac 180aaccagaagt tcaagggcag
ggtgaccatc accgccgaca aaagcaccag caccgcctac 240atggaactga
gcagcctgag gagcgaggac accgccgtgt actactgcgc caacggctac
300gagttcgtgt attggggcca gggcacacta gtgaccgtgt ccagc
34583115PRTArtificial SequenceHumansed antibody sequence 83Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20
25 30 Tyr Met Lys Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Ile 35 40 45 Gly Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Gly Tyr Glu Phe Val
Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115
84345DNAArtificial SequenceHumansed antibody sequence 84caggtgcagc
tggtgcagag cggcgccgaa gtgaagaagc ccggctccag cgtgaaggtg 60agctgcaagg
ctagcggcta caccttcacc gactactaca tgaagtgggt gaggcaggcc
120cccggccagg gactggagtg gataggcgag atctacccca acaacggggg
catcacctac 180aaccagaagt tcaagggcag ggcgaccctc accgtcgaca
aaagcaccag caccgcctac 240atggaactga gcagcctgag gagcgaggac
accgccgtgt actactgcgc caacggctac 300gagttcgtgt attggggcca
gggcacacta gtgaccgtgt ccagc 34585115PRTArtificial SequenceHumansed
antibody sequence 85Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys
Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly
Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys Trp Val Arg Gln Ala
Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Glu Ile Tyr Pro Asn
Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Val
Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu
Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Ala Asp Gly Tyr Glu Phe Val Tyr Trp Gly Gln Gly Thr Leu Val Thr 100
105 110 Val Ser Ser 115 86345DNAArtificial SequenceHumansed
antibody sequence 86caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc
ccggctccag cgtgaaggtg 60agctgcaagg ctagcggcta caccttcacc gactactaca
tgaagtgggt gaggcaggcc 120cccggccagg gactggagtg gatgggcgag
atctacccca acaacggggg catcacctac 180aaccagaagt tcaagggcag
ggtgaccatc accgccgaca aaagcaccag caccgcctac 240atggaactga
gcagcctgag gagcgaggac accgccgtgt actactgcgc cgacggctac
300gagttcgtgt attggggcca gggcacacta gtgaccgtgt ccagc
34587115PRTArtificial SequenceHumansed antibody sequence 87Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20
25 30 Tyr Met Lys Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Ile 35 40 45 Gly Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn
Gln Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser
Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Gly Tyr Glu Phe Asp
Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 115
88345DNAArtificial SequenceHumansed antibody sequence 88caggtgcagc
tggtgcagag cggcgccgaa gtgaagaagc ccggctccag cgtgaaggtg 60agctgcaagg
ctagcggcta caccttcacc gactactaca tgaagtgggt gaggcaggcc
120cccggccagg gactggagtg gataggcgag atctacccca acaacggggg
catcacctac 180aaccagaagt tcaagggcag ggcgaccctc accgtcgaca
aaagcaccag caccgcctac 240atggaactga gcagcctgag gagcgaggac
accgccgtgt actactgcgc caacggctac 300gagttcgact attggggcca
gggcacacta gtgaccgtgt ccagc 34589108PRTArtificial SequenceHumansed
antibody sequence 89Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly1 5 10 15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser
Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu
His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80 Glu Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Lys Leu Pro Trp 85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105
90324DNAArtificial SequenceHumansed antibody sequence 90gacatccaga
tgacccagag cccctcaagc ctgagcgcca gcgtgggcga cagggtgact 60atcacctgca
gcgcctccca gggcatcagc aactacctga actggtacca gcagaagccc
120ggcaaggccc ctaagctgct gatctactac accagcagcc tgcacagcgg
cgtgcccagc 180aggttctccg gcagcggcag cggaaccgac ttcaccctga
ccattagcag cctccagccc 240gaggacttcg ccacctacta ctgccagcag
tacagcaagc tgccctggac cttcggccag 300ggcaccaaac tggagatcaa gcgt
32491108PRTArtificial SequenceHumansed antibody sequence 91Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr 20
25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser 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 Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Tyr Ser Lys Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys
Leu Glu Ile Lys Arg 100 105 92324DNAArtificial SequenceHumansed
antibody sequence 92gacatccaga tgacccagag cccctcaagc ctgagcgcca
gcgtgggcga cagggtgact 60atcacctgca gcgcctccca gggcatcagc aactacctga
actggtacca gcagaagccc 120ggcaaggccc ctaagctgct gatctactac
accagcagcc tgcacagcgg cgtgcccagc 180aggttctccg gcagcggcag
cggaaccgac tacaccctga ccattagcag cctccagccc 240gaggacttcg
ccacctacta ctgccagcag tacagcaagc tgccctggac cttcggccag
300ggcaccaaac tggagatcaa gcgt 324935PRTmus musculus 93Asp Tyr Tyr
Met Lys1 5 9417PRTmus musculus 94Glu Ile Tyr Pro Asn Asn Gly Gly
Ile Thr Tyr Asn Gln Lys Phe Lys1 5 10 15 Gly956PRTmus musculus
95Gly Tyr Glu Phe Val Tyr1 5 9611PRTmus musculus 96Ser Ala Ser Gln
Gly Ile Ser Asn Tyr Leu Asn1 5 10 977PRTmus musculus 97Tyr Thr Ser
Ser Leu His Ser1 5 989PRTmus musculus 98Gln Gln Tyr Ser Lys Leu Pro
Trp Thr1 5 996PRTArtificial SequenceHumansed antibody sequence
99Gly Tyr Glu Phe Asp Tyr1 5 100445PRTArtificial SequenceHumansed
antibody sequence 100Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val
Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser
Gly Gly Thr Phe Ser Asp Tyr 20 25 30 Tyr Met Lys Trp Val Arg Gln
Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Glu Ile Tyr Pro
Asn Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg
Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met
Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Gly Tyr Glu Phe Val Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110 Val Ser Ser 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 Ala145 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 Asp Lys Thr His Thr Cys 210 215
220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
Leu225 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 Lys305 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 Gly385 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
1011335DNAArtificial SequenceHumansed antibody sequence
101caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc ccggctccag
cgtgaaggtg 60agctgcaagg ctagcggcgg caccttcagc gactactaca tgaagtgggt
gaggcaggcc 120cccggccagg gactggagtg gatgggcgag atctacccca
acaacggggg catcacctac 180aaccagaagt tcaagggcag ggtgaccatc
accgccgaca aaagcaccag caccgcctac 240atggaactga gcagcctgag
gagcgaggac accgccgtgt actactgcgc caggggctac 300gagttcgtgt
attggggcca gggcacacta gtgaccgtgt ccagcgccag caccaagggc
360cccagcgtgt tccccctggc ccccagcagc aagagcacca gcggcggcac
agccgccctg 420ggctgcctgg tgaaggacta cttccccgaa ccggtgaccg
tgtcctggaa cagcggagcc 480ctgaccagcg gcgtgcacac cttccccgcc
gtgctgcaga gcagcggcct gtacagcctg 540agcagcgtgg tgaccgtgcc
cagcagcagc ctgggcaccc agacctacat ctgtaacgtg 600aaccacaagc
ccagcaacac caaggtggac aagaaggtgg agcccaagag ctgtgacaag
660acccacacct gccccccctg ccctgccccc gagctgctgg gaggccccag
cgtgttcctg 720ttccccccca agcctaagga caccctgatg atcagcagaa
cccccgaggt gacctgtgtg 780gtggtggatg tgagccacga ggaccctgag
gtgaagttca actggtacgt ggacggcgtg 840gaggtgcaca atgccaagac
caagcccagg gaggagcagt acaacagcac ctaccgggtg 900gtgtccgtgc
tgaccgtgct gcaccaggat tggctgaacg gcaaggagta caagtgtaag
960gtgtccaaca aggccctgcc tgcccctatc gagaaaacca tcagcaaggc
caagggccag 1020cccagagagc cccaggtgta caccctgccc cctagcagag
atgagctgac caagaaccag 1080gtgtccctga cctgcctggt gaagggcttc
taccccagcg acatcgccgt ggagtgggag 1140agcaacggcc agcccgagaa
caactacaag accacccccc ctgtgctgga cagcgatggc 1200agcttcttcc
tgtacagcaa gctgaccgtg gacaagagca gatggcagca gggcaacgtg
1260ttcagctgct ccgtgatgca cgaggccctg cacaatcact acacccagaa
gagcctgagc 1320ctgtcccctg gcaag 1335102445PRTArtificial
SequenceHumansed antibody sequence 102Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Gly Tyr Glu Phe Val Tyr Trp Gly Gln
Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 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 Ala145 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 Asp Lys Thr
His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu225 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 Lys305
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 Gly385 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
1031335DNAArtificial SequenceHumansed antibody sequence
103caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc ccggctccag
cgtgaaggtg 60agctgcaagg ctagcggcta caccttcacc gactactaca tgaagtgggt
gaggcaggcc 120cccggccagg gactggagtg gatgggcgag atctacccca
acaacggggg catcacctac 180aaccagaagt tcaagggcag ggtgaccatc
accgccgaca aaagcaccag caccgcctac 240atggaactga gcagcctgag
gagcgaggac accgccgtgt actactgcgc caggggctac 300gagttcgtgt
attggggcca gggcacacta gtgaccgtgt ccagcgccag caccaagggc
360cccagcgtgt tccccctggc ccccagcagc aagagcacca gcggcggcac
agccgccctg 420ggctgcctgg tgaaggacta cttccccgaa ccggtgaccg
tgtcctggaa cagcggagcc 480ctgaccagcg gcgtgcacac cttccccgcc
gtgctgcaga gcagcggcct gtacagcctg 540agcagcgtgg tgaccgtgcc
cagcagcagc ctgggcaccc agacctacat ctgtaacgtg 600aaccacaagc
ccagcaacac caaggtggac aagaaggtgg agcccaagag ctgtgacaag
660acccacacct gccccccctg ccctgccccc gagctgctgg gaggccccag
cgtgttcctg 720ttccccccca agcctaagga caccctgatg atcagcagaa
cccccgaggt gacctgtgtg 780gtggtggatg tgagccacga ggaccctgag
gtgaagttca actggtacgt ggacggcgtg 840gaggtgcaca atgccaagac
caagcccagg gaggagcagt acaacagcac ctaccgggtg 900gtgtccgtgc
tgaccgtgct gcaccaggat tggctgaacg gcaaggagta caagtgtaag
960gtgtccaaca aggccctgcc tgcccctatc gagaaaacca tcagcaaggc
caagggccag 1020cccagagagc cccaggtgta caccctgccc cctagcagag
atgagctgac caagaaccag 1080gtgtccctga cctgcctggt gaagggcttc
taccccagcg acatcgccgt ggagtgggag 1140agcaacggcc agcccgagaa
caactacaag accacccccc ctgtgctgga cagcgatggc 1200agcttcttcc
tgtacagcaa gctgaccgtg gacaagagca gatggcagca gggcaacgtg
1260ttcagctgct ccgtgatgca cgaggccctg cacaatcact acacccagaa
gagcctgagc 1320ctgtcccctg gcaag 1335104445PRTArtificial
SequenceHumansed antibody sequence 104Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Asn Gly Tyr Glu Phe Val Tyr Trp Gly Gln
Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 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 Ala145 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 Asp Lys Thr
His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu225 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 Lys305
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 Gly385 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
1051335DNAArtificial SequenceHumansed antibody sequence
105caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc ccggctccag
cgtgaaggtg 60agctgcaagg ctagcggcta caccttcacc gactactaca tgaagtgggt
gaggcaggcc 120cccggccagg gactggagtg gatgggcgag atctacccca
acaacggggg catcacctac 180aaccagaagt tcaagggcag ggtgaccatc
accgccgaca aaagcaccag caccgcctac 240atggaactga gcagcctgag
gagcgaggac accgccgtgt actactgcgc caacggctac 300gagttcgtgt
attggggcca gggcacacta gtgaccgtgt ccagcgccag caccaagggc
360cccagcgtgt tccccctggc ccccagcagc aagagcacca gcggcggcac
agccgccctg 420ggctgcctgg tgaaggacta cttccccgaa ccggtgaccg
tgtcctggaa cagcggagcc 480ctgaccagcg gcgtgcacac cttccccgcc
gtgctgcaga gcagcggcct gtacagcctg 540agcagcgtgg tgaccgtgcc
cagcagcagc ctgggcaccc agacctacat ctgtaacgtg 600aaccacaagc
ccagcaacac caaggtggac aagaaggtgg agcccaagag ctgtgacaag
660acccacacct gccccccctg ccctgccccc gagctgctgg gaggccccag
cgtgttcctg 720ttccccccca agcctaagga caccctgatg atcagcagaa
cccccgaggt gacctgtgtg 780gtggtggatg tgagccacga ggaccctgag
gtgaagttca actggtacgt ggacggcgtg 840gaggtgcaca atgccaagac
caagcccagg gaggagcagt acaacagcac ctaccgggtg 900gtgtccgtgc
tgaccgtgct gcaccaggat tggctgaacg gcaaggagta caagtgtaag
960gtgtccaaca aggccctgcc tgcccctatc gagaaaacca tcagcaaggc
caagggccag 1020cccagagagc cccaggtgta caccctgccc cctagcagag
atgagctgac caagaaccag 1080gtgtccctga cctgcctggt gaagggcttc
taccccagcg acatcgccgt ggagtgggag 1140agcaacggcc agcccgagaa
caactacaag accacccccc ctgtgctgga cagcgatggc 1200agcttcttcc
tgtacagcaa gctgaccgtg gacaagagca gatggcagca gggcaacgtg
1260ttcagctgct ccgtgatgca cgaggccctg cacaatcact acacccagaa
gagcctgagc 1320ctgtcccctg gcaag 1335106445PRTArtificial
SequenceHumansed antibody sequence 106Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly
Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Asn Gly Tyr Glu Phe Val Tyr Trp Gly Gln
Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 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 Ala145 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 Asp Lys Thr
His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu225 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 Lys305
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 Gly385 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
1071335DNAArtificial SequenceHumansed antibody sequence
107caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc ccggctccag
cgtgaaggtg 60agctgcaagg ctagcggcta caccttcacc gactactaca tgaagtgggt
gaggcaggcc 120cccggccagg gactggagtg gataggcgag atctacccca
acaacggggg catcacctac 180aaccagaagt tcaagggcag ggcgaccctc
accgtcgaca aaagcaccag caccgcctac 240atggaactga gcagcctgag
gagcgaggac accgccgtgt actactgcgc caacggctac 300gagttcgtgt
attggggcca gggcacacta gtgaccgtgt ccagcgccag caccaagggc
360cccagcgtgt tccccctggc ccccagcagc aagagcacca gcggcggcac
agccgccctg 420ggctgcctgg tgaaggacta cttccccgaa ccggtgaccg
tgtcctggaa cagcggagcc 480ctgaccagcg gcgtgcacac cttccccgcc
gtgctgcaga gcagcggcct gtacagcctg 540agcagcgtgg tgaccgtgcc
cagcagcagc ctgggcaccc agacctacat ctgtaacgtg 600aaccacaagc
ccagcaacac caaggtggac aagaaggtgg agcccaagag ctgtgacaag
660acccacacct gccccccctg ccctgccccc gagctgctgg gaggccccag
cgtgttcctg 720ttccccccca agcctaagga caccctgatg atcagcagaa
cccccgaggt gacctgtgtg 780gtggtggatg tgagccacga ggaccctgag
gtgaagttca actggtacgt ggacggcgtg 840gaggtgcaca atgccaagac
caagcccagg gaggagcagt acaacagcac ctaccgggtg 900gtgtccgtgc
tgaccgtgct gcaccaggat tggctgaacg gcaaggagta caagtgtaag
960gtgtccaaca aggccctgcc tgcccctatc gagaaaacca tcagcaaggc
caagggccag 1020cccagagagc cccaggtgta caccctgccc cctagcagag
atgagctgac caagaaccag 1080gtgtccctga cctgcctggt gaagggcttc
taccccagcg acatcgccgt ggagtgggag 1140agcaacggcc agcccgagaa
caactacaag accacccccc ctgtgctgga cagcgatggc 1200agcttcttcc
tgtacagcaa gctgaccgtg gacaagagca gatggcagca gggcaacgtg
1260ttcagctgct ccgtgatgca cgaggccctg cacaatcact acacccagaa
gagcctgagc 1320ctgtcccctg gcaag 1335108445PRTArtificial
SequenceHumansed antibody sequence 108Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly
Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95 Ala Asp Gly Tyr Glu Phe Val Tyr Trp Gly Gln
Gly Thr Leu Val Thr 100 105 110 Val Ser Ser 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 Ala145 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 Asp Lys Thr
His Thr Cys 210 215 220 Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
Pro Ser Val Phe Leu225 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 Lys305
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 Gly385 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
1091335DNAArtificial SequenceHumansed antibody sequence
109caggtgcagc tggtgcagag cggcgccgaa gtgaagaagc ccggctccag
cgtgaaggtg 60agctgcaagg ctagcggcta caccttcacc gactactaca tgaagtgggt
gaggcaggcc 120cccggccagg gactggagtg gatgggcgag atctacccca
acaacggggg catcacctac 180aaccagaagt tcaagggcag ggtgaccatc
accgccgaca aaagcaccag caccgcctac 240atggaactga gcagcctgag
gagcgaggac accgccgtgt actactgcgc cgacggctac 300gagttcgtgt
attggggcca gggcacacta gtgaccgtgt ccagcgccag caccaagggc
360cccagcgtgt tccccctggc ccccagcagc aagagcacca gcggcggcac
agccgccctg 420ggctgcctgg tgaaggacta cttccccgaa ccggtgaccg
tgtcctggaa cagcggagcc 480ctgaccagcg gcgtgcacac cttccccgcc
gtgctgcaga gcagcggcct gtacagcctg 540agcagcgtgg tgaccgtgcc
cagcagcagc ctgggcaccc agacctacat ctgtaacgtg 600aaccacaagc
ccagcaacac caaggtggac aagaaggtgg agcccaagag ctgtgacaag
660acccacacct gccccccctg ccctgccccc gagctgctgg gaggccccag
cgtgttcctg 720ttccccccca agcctaagga caccctgatg atcagcagaa
cccccgaggt gacctgtgtg 780gtggtggatg tgagccacga ggaccctgag
gtgaagttca actggtacgt ggacggcgtg 840gaggtgcaca atgccaagac
caagcccagg gaggagcagt acaacagcac ctaccgggtg 900gtgtccgtgc
tgaccgtgct gcaccaggat tggctgaacg gcaaggagta caagtgtaag
960gtgtccaaca aggccctgcc tgcccctatc gagaaaacca tcagcaaggc
caagggccag 1020cccagagagc cccaggtgta caccctgccc cctagcagag
atgagctgac caagaaccag 1080gtgtccctga cctgcctggt gaagggcttc
taccccagcg acatcgccgt ggagtgggag 1140agcaacggcc agcccgagaa
caactacaag accacccccc ctgtgctgga cagcgatggc 1200agcttcttcc
tgtacagcaa gctgaccgtg gacaagagca gatggcagca gggcaacgtg
1260ttcagctgct ccgtgatgca cgaggccctg cacaatcact acacccagaa
gagcctgagc 1320ctgtcccctg gcaag 1335110445PRTArtificial
SequenceHumansed antibody sequence 110Gln Val Gln Leu Val Gln Ser
Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15 Ser Val Lys Val Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met Lys
Trp Val Arg Gln Ala Pro Gly Gln Gly Leu
Glu Trp Ile 35 40 45 Gly Glu Ile Tyr Pro Asn Asn Gly Gly Ile Thr
Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Arg Ala Thr Leu Thr Val Asp
Lys Ser Thr Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Gly Tyr Glu
Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr 100 105 110 Val Ser Ser
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
Ala145 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 Asp Lys Thr His Thr Cys 210 215 220 Pro Pro Cys Pro
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu225 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 Lys305 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
Gly385 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 1111335DNAArtificial
SequenceHumansed antibody sequence 111caggtgcagc tggtgcagag
cggcgccgaa gtgaagaagc ccggctccag cgtgaaggtg 60agctgcaagg ctagcggcta
caccttcacc gactactaca tgaagtgggt gaggcaggcc 120cccggccagg
gactggagtg gataggcgag atctacccca acaacggggg catcacctac
180aaccagaagt tcaagggcag ggcgaccctc accgtcgaca aaagcaccag
caccgcctac 240atggaactga gcagcctgag gagcgaggac accgccgtgt
actactgcgc caacggctac 300gagttcgact attggggcca gggcacacta
gtgaccgtgt ccagcgccag caccaagggc 360cccagcgtgt tccccctggc
ccccagcagc aagagcacca gcggcggcac agccgccctg 420ggctgcctgg
tgaaggacta cttccccgaa ccggtgaccg tgtcctggaa cagcggagcc
480ctgaccagcg gcgtgcacac cttccccgcc gtgctgcaga gcagcggcct
gtacagcctg 540agcagcgtgg tgaccgtgcc cagcagcagc ctgggcaccc
agacctacat ctgtaacgtg 600aaccacaagc ccagcaacac caaggtggac
aagaaggtgg agcccaagag ctgtgacaag 660acccacacct gccccccctg
ccctgccccc gagctgctgg gaggccccag cgtgttcctg 720ttccccccca
agcctaagga caccctgatg atcagcagaa cccccgaggt gacctgtgtg
780gtggtggatg tgagccacga ggaccctgag gtgaagttca actggtacgt
ggacggcgtg 840gaggtgcaca atgccaagac caagcccagg gaggagcagt
acaacagcac ctaccgggtg 900gtgtccgtgc tgaccgtgct gcaccaggat
tggctgaacg gcaaggagta caagtgtaag 960gtgtccaaca aggccctgcc
tgcccctatc gagaaaacca tcagcaaggc caagggccag 1020cccagagagc
cccaggtgta caccctgccc cctagcagag atgagctgac caagaaccag
1080gtgtccctga cctgcctggt gaagggcttc taccccagcg acatcgccgt
ggagtgggag 1140agcaacggcc agcccgagaa caactacaag accacccccc
ctgtgctgga cagcgatggc 1200agcttcttcc tgtacagcaa gctgaccgtg
gacaagagca gatggcagca gggcaacgtg 1260ttcagctgct ccgtgatgca
cgaggccctg cacaatcact acacccagaa gagcctgagc 1320ctgtcccctg gcaag
1335112214PRTArtificial SequenceHumansed antibody sequence 112Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15 Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr
20 25 30 Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu
Leu Ile 35 40 45 Tyr Tyr Thr Ser Ser Leu His Ser Gly Val Pro Ser
Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser Ser Leu Gln Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Tyr Ser Lys Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145
150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser
Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210
113642DNAArtificial SequenceHumansed antibody sequence
113gacatccaga tgacccagag cccctcaagc ctgagcgcca gcgtgggcga
cagggtgact 60atcacctgca gcgcctccca gggcatcagc aactacctga actggtacca
gcagaagccc 120ggcaaggccc ctaagctgct gatctactac accagcagcc
tgcacagcgg cgtgcccagc 180aggttctccg gcagcggcag cggaaccgac
ttcaccctga ccattagcag cctccagccc 240gaggacttcg ccacctacta
ctgccagcag tacagcaagc tgccctggac cttcggccag 300ggcaccaaac
tggagatcaa gcgtacggtg gccgccccca gcgtgttcat cttccccccc
360agcgatgagc agctgaagag cggcaccgcc agcgtggtgt gtctgctgaa
caacttctac 420ccccgggagg ccaaggtgca gtggaaggtg gacaatgccc
tgcagagcgg caacagccag 480gagagcgtga ccgagcagga cagcaaggac
tccacctaca gcctgagcag caccctgacc 540ctgagcaagg ccgactacga
gaagcacaag gtgtacgcct gtgaggtgac ccaccagggc 600ctgtccagcc
ccgtgaccaa gagcttcaac cggggcgagt gc 642114214PRTArtificial
SequenceHumansed antibody sequence 114Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15 Asp Arg Val Thr Ile
Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr 20 25 30 Leu Asn Trp
Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr
Tyr Thr Ser Ser Leu His Ser 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
Pro65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Lys
Leu Pro Trp 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser
Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu
Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys
Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160 Glu Ser
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180
185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys
Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 115642DNAArtificial
SequenceHumansed antibody sequence 115gacatccaga tgacccagag
cccctcaagc ctgagcgcca gcgtgggcga cagggtgact 60atcacctgca gcgcctccca
gggcatcagc aactacctga actggtacca gcagaagccc 120ggcaaggccc
ctaagctgct gatctactac accagcagcc tgcacagcgg cgtgcccagc
180aggttctccg gcagcggcag cggaaccgac tacaccctga ccattagcag
cctccagccc 240gaggacttcg ccacctacta ctgccagcag tacagcaagc
tgccctggac cttcggccag 300ggcaccaaac tggagatcaa gcgtacggtg
gccgccccca gcgtgttcat cttccccccc 360agcgatgagc agctgaagag
cggcaccgcc agcgtggtgt gtctgctgaa caacttctac 420ccccgggagg
ccaaggtgca gtggaaggtg gacaatgccc tgcagagcgg caacagccag
480gagagcgtga ccgagcagga cagcaaggac tccacctaca gcctgagcag
caccctgacc 540ctgagcaagg ccgactacga gaagcacaag gtgtacgcct
gtgaggtgac ccaccagggc 600ctgtccagcc ccgtgaccaa gagcttcaac
cggggcgagt gc 642116118PRTmus musculus 116Glu Val Gln Leu Gln Gln
Ser Gly Pro Val Leu Val Lys Pro Gly Ala1 5 10 15 Ser Val Lys Met
Ser Cys Glu Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Tyr Met
Asn Trp Val Lys Gln Ser His Gly Lys Thr Leu Glu Trp Ile 35 40 45
Gly Val Ile Asn Pro Tyr Asn Gly Gly Thr Asp Tyr Asn Gln Lys Phe 50
55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Asn Ser Leu Thr Ser Glu Asp Ser Ala Val
Tyr Tyr Cys 85 90 95 Ala Arg Ser Val Tyr Asp Tyr Pro Phe Asp Tyr
Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115
117354DNAmus musculus 117gaggtgcagc tgcagcagag cggccccgtg
ctggtgaagc ctggagccag cgtgaaaatg 60agctgcgaag ccagcggcta caccttcacc
gactactaca tgaactgggt gaagcagagc 120cacggcaaga ccctggagtg
gatcggcgtg atcaacccct acaacggggg caccgactac 180aaccagaagt
tcaagggcaa ggccactctg accgtggaca agagctccag caccgcctac
240atggaactga acagcctcac ctctgaggac agcgccgtct attactgcgc
caggagcgtg 300tacgactacc ccttcgacta ctggggccag ggcacactag
tgaccgtgtc cagc 354118448PRTArtificial SequenceChimeric antibody
sequence 118Glu Val Gln Leu Gln Gln Ser Gly Pro Val Leu Val Lys Pro
Gly Ala1 5 10 15 Ser Val Lys Met Ser Cys Glu Ala Ser Gly Tyr Thr
Phe Thr Asp Tyr 20 25 30 Tyr Met Asn Trp Val Lys Gln Ser His Gly
Lys Thr Leu Glu Trp Ile 35 40 45 Gly Val Ile Asn Pro Tyr Asn Gly
Gly Thr Asp Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu
Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Asn
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg
Ser Val Tyr Asp Tyr Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115
120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser225 230 235
240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr305 310 315 320 Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360
365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
1191344DNAArtificial SequenceChimeric antibody sequence
119gaggtgcagc tgcagcagag cggccccgtg ctggtgaagc ctggagccag
cgtgaaaatg 60agctgcgaag ccagcggcta caccttcacc gactactaca tgaactgggt
gaagcagagc 120cacggcaaga ccctggagtg gatcggcgtg atcaacccct
acaacggggg caccgactac 180aaccagaagt tcaagggcaa ggccactctg
accgtggaca agagctccag caccgcctac 240atggaactga acagcctcac
ctctgaggac agcgccgtct attactgcgc caggagcgtg 300tacgactacc
ccttcgacta ctggggccag ggcacactag tgaccgtgtc cagcgccagc
360accaagggcc ccagcgtgtt ccccctggcc cccagcagca agagcaccag
cggcggcaca 420gccgccctgg gctgcctggt gaaggactac ttccccgaac
cggtgaccgt gtcctggaac 480agcggagccc tgaccagcgg cgtgcacacc
ttccccgccg tgctgcagag cagcggcctg 540tacagcctga gcagcgtggt
gaccgtgccc agcagcagcc tgggcaccca gacctacatc 600tgtaacgtga
accacaagcc cagcaacacc aaggtggaca agaaggtgga gcccaagagc
660tgtgacaaga cccacacctg ccccccctgc cctgcccccg agctgctggg
aggccccagc 720gtgttcctgt tcccccccaa gcctaaggac accctgatga
tcagcagaac ccccgaggtg 780acctgtgtgg tggtggatgt gagccacgag
gaccctgagg tgaagttcaa ctggtacgtg 840gacggcgtgg aggtgcacaa
tgccaagacc aagcccaggg aggagcagta caacagcacc 900taccgggtgg
tgtccgtgct gaccgtgctg caccaggatt ggctgaacgg caaggagtac
960aagtgtaagg tgtccaacaa ggccctgcct gcccctatcg agaaaaccat
cagcaaggcc 1020aagggccagc ccagagagcc ccaggtgtac accctgcccc
ctagcagaga tgagctgacc 1080aagaaccagg tgtccctgac ctgcctggtg
aagggcttct accccagcga catcgccgtg 1140gagtgggaga gcaacggcca
gcccgagaac aactacaaga ccaccccccc tgtgctggac 1200agcgatggca
gcttcttcct gtacagcaag ctgaccgtgg acaagagcag atggcagcag
1260ggcaacgtgt tcagctgctc cgtgatgcac gaggccctgc acaatcacta
cacccagaag 1320agcctgagcc tgtcccctgg caag 1344120111PRTmus musculus
120Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1
5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Ile
His 20 25 30 Gly Thr His Leu Met His Trp Tyr Gln Gln Lys Pro Gly
Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu
Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Glu Thr
Asp Phe Thr Leu Asn Ile His65 70 75 80 Pro Val Glu Glu Glu Asp Ala
Ala Thr Tyr Phe Cys Gln Gln Ser Ile 85 90 95 Glu Asp Pro Arg Thr
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 110 121336DNAmus
musculus 121gacatcgtcc tgacccagag ccccgccagc ctggccgtga gcctgggcca
gagggccaca 60atcagctgca gggcctctga gtccgtgagc atccacggca cccacctgat
gcactggtat 120cagcagaagc ccggccagcc tcccaagctg ctgatctacg
ccgccagcaa cctggagagc 180ggcgtgcccg ctaggttcag cggaagcggc
agcgagaccg acttcaccct gaacatccac 240cccgtggagg aggaagacgc
cgccacctac ttctgccagc agagcatcga ggaccccagg 300accttcggcg
ggggcaccaa
gctcgagatt aagcgt 336122237PRTArtificial SequenceChimeric antibody
sequence 122Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala
Thr Gly1 5 10 15 Val His Ser Asp Ile Val Leu Thr Gln Ser Pro Ala
Ser Leu Ala Val 20 25 30 Ser Leu Gly Gln Arg Ala Thr Ile Ser Cys
Arg Ala Ser Glu Ser Val 35 40 45 Ser Ile His Gly Thr His Leu Met
His Trp Tyr Gln Gln Lys Pro Gly 50 55 60 Gln Pro Pro Lys Leu Leu
Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly65 70 75 80 Val Pro Ala Arg
Phe Ser Gly Ser Gly Ser Glu Thr Asp Phe Thr Leu 85 90 95 Asn Ile
His Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Phe Cys Gln 100 105 110
Gln Ser Ile Glu Asp Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu 115
120 125 Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro
Ser 130 135 140 Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys
Leu Leu Asn145 150 155 160 Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln
Trp Lys Val Asp Asn Ala 165 170 175 Leu Gln Ser Gly Asn Ser Gln Glu
Ser Val Thr Glu Gln Asp Ser Lys 180 185 190 Asp Ser Thr Tyr Ser Leu
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp 195 200 205 Tyr Glu Lys His
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu 210 215 220 Ser Ser
Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230 235
123711DNAArtificial SequenceChimeric antibody sequence
123atgggctggt cctgcatcat cctgtttctg gtggccaccg ccaccggcgt
gcacagcgac 60atcgtcctga cccagagccc cgccagcctg gccgtgagcc tgggccagag
ggccacaatc 120agctgcaggg cctctgagtc cgtgagcatc cacggcaccc
acctgatgca ctggtatcag 180cagaagcccg gccagcctcc caagctgctg
atctacgccg ccagcaacct ggagagcggc 240gtgcccgcta ggttcagcgg
aagcggcagc gagaccgact tcaccctgaa catccacccc 300gtggaggagg
aagacgccgc cacctacttc tgccagcaga gcatcgagga ccccaggacc
360ttcggcgggg gcaccaagct cgagattaag cgtacggtgg ccgcccccag
cgtgttcatc 420ttccccccca gcgatgagca gctgaagagc ggcaccgcca
gcgtggtgtg tctgctgaac 480aacttctacc cccgggaggc caaggtgcag
tggaaggtgg acaatgccct gcagagcggc 540aacagccagg agagcgtgac
cgagcaggac agcaaggact ccacctacag cctgagcagc 600accctgaccc
tgagcaaggc cgactacgag aagcacaagg tgtacgcctg tgaggtgacc
660caccagggcc tgtccagccc cgtgaccaag agcttcaacc ggggcgagtg c
711124119PRTmus musculus 124Glu Val Gln Leu Gln Gln Ser Gly Pro Glu
Leu Val Lys Pro Gly Thr1 5 10 15 Ser Val Lys Ile Pro Cys Lys Thr
Ser Gly Tyr Ile Phe Thr Asp Tyr 20 25 30 Ser Ile Asp Trp Val Lys
Gln Ser His Gly Lys Ser Leu Glu Trp Ile 35 40 45 Gly Asp Ile Asp
Pro Asn Tyr Gly Asp Pro Ile Tyr Asn His Lys Phe 50 55 60 Lys Gly
Lys Ala Thr Leu Thr Val Asp Arg Ser Ser Ser Thr Ala Tyr65 70 75 80
Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Thr Ala Val Tyr Phe Cys 85
90 95 Ala Arg Arg Ala Thr Gly Thr Asp Trp Phe Ala Phe Trp Gly Gln
Gly 100 105 110 Thr Leu Val Thr Val Ser Ser 115 125357DNAmus
musculus 125gaggtgcagc tgcagcagag cggccccgag ctggtgaaac ccggcaccag
cgtgaagatc 60ccctgcaaga cctctggcta catcttcacc gactacagca tcgactgggt
gaagcagagc 120cacggcaagt ctctggagtg gattggggac atcgacccca
actacggcga ccccatctac 180aaccacaagt tcaagggcaa ggccaccctg
accgtggaca ggagcagcag caccgcctac 240atggaactca ggagcctgac
cagcgaggac accgccgtgt atttttgcgc caggagggcc 300accggcactg
attggttcgc cttctggggc cagggcacac tagtgaccgt gtccagc
357126449PRTArtificial SequenceChimeric antibody sequence 126Glu
Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Thr1 5 10
15 Ser Val Lys Ile Pro Cys Lys Thr Ser Gly Tyr Ile Phe Thr Asp Tyr
20 25 30 Ser Ile Asp Trp Val Lys Gln Ser His Gly Lys Ser Leu Glu
Trp Ile 35 40 45 Gly Asp Ile Asp Pro Asn Tyr Gly Asp Pro Ile Tyr
Asn His Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu Thr Val Asp Arg
Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser
Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Arg Arg Ala Thr Gly
Thr Asp Trp Phe Ala Phe Trp Gly Gln Gly 100 105 110 Thr Leu Val Thr
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu
Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp145
150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala
Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys
Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro225 230 235 240 Ser Val Phe
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265
270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn
275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
Asn Gly Lys Glu305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala
Leu Pro Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly
Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg
Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu385 390
395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
Ser Leu Ser Pro Gly 435 440 445 Lys 1271347DNAArtificial
SequenceChimeric antibody sequence 127gaggtgcagc tgcagcagag
cggccccgag ctggtgaaac ccggcaccag cgtgaagatc 60ccctgcaaga cctctggcta
catcttcacc gactacagca tcgactgggt gaagcagagc 120cacggcaagt
ctctggagtg gattggggac atcgacccca actacggcga ccccatctac
180aaccacaagt tcaagggcaa ggccaccctg accgtggaca ggagcagcag
caccgcctac 240atggaactca ggagcctgac cagcgaggac accgccgtgt
atttttgcgc caggagggcc 300accggcactg attggttcgc cttctggggc
cagggcacac tagtgaccgt gtccagcgcc 360agcaccaagg gccccagcgt
gttccccctg gcccccagca gcaagagcac cagcggcggc 420acagccgccc
tgggctgcct ggtgaaggac tacttccccg aaccggtgac cgtgtcctgg
480aacagcggag ccctgaccag cggcgtgcac accttccccg ccgtgctgca
gagcagcggc 540ctgtacagcc tgagcagcgt ggtgaccgtg cccagcagca
gcctgggcac ccagacctac 600atctgtaacg tgaaccacaa gcccagcaac
accaaggtgg acaagaaggt ggagcccaag 660agctgtgaca agacccacac
ctgccccccc tgccctgccc ccgagctgct gggaggcccc 720agcgtgttcc
tgttcccccc caagcctaag gacaccctga tgatcagcag aacccccgag
780gtgacctgtg tggtggtgga tgtgagccac gaggaccctg aggtgaagtt
caactggtac 840gtggacggcg tggaggtgca caatgccaag accaagccca
gggaggagca gtacaacagc 900acctaccggg tggtgtccgt gctgaccgtg
ctgcaccagg attggctgaa cggcaaggag 960tacaagtgta aggtgtccaa
caaggccctg cctgccccta tcgagaaaac catcagcaag 1020gccaagggcc
agcccagaga gccccaggtg tacaccctgc cccctagcag agatgagctg
1080accaagaacc aggtgtccct gacctgcctg gtgaagggct tctaccccag
cgacatcgcc 1140gtggagtggg agagcaacgg ccagcccgag aacaactaca
agaccacccc ccctgtgctg 1200gacagcgatg gcagcttctt cctgtacagc
aagctgaccg tggacaagag cagatggcag 1260cagggcaacg tgttcagctg
ctccgtgatg cacgaggccc tgcacaatca ctacacccag 1320aagagcctga
gcctgtcccc tggcaag 1347128108PRTmus musculus 128Asp Ile Gln Met Thr
Gln Ser Pro Ala Ser Leu Ser Val Ser Val Gly1 5 10 15 Glu Thr Val
Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Asn Asn 20 25 30 Leu
Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu Leu Val 35 40
45 Tyr Ala Ala Thr Ile Leu Ala Asp Gly Val Pro Ser Arg Phe Ser Gly
50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys Ile Asn Ser Leu
Gln Ser65 70 75 80 Gly Asp Phe Gly Thr Tyr Tyr Cys Gln His Phe Trp
Gly Thr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
Lys Arg 100 105 129324DNAmus musculus 129gacatccaga tgacccagag
ccccgctagc ctcagcgtgt ccgtcggcga gaccgtgacc 60atcacctgca gggccagcga
gaacatctac aacaacctgg cctggtatca gcagaagcag 120ggcaaaagcc
cccagctgct ggtgtacgcc gccaccattc tggccgacgg cgtgcccagc
180aggttctctg gaagcggcag cggcacccag tacagcctga agatcaacag
cctgcagagc 240ggggacttcg gcacctacta ctgccagcac ttctggggca
ctcccctgac cttcggagcc 300ggcaccaagc tggagctgaa gcgt
324130214PRTArtificial SequenceChimeric antibody sequence 130Asp
Ile Gln Met Thr Gln Ser Pro Ala Ser Leu Ser Val Ser Val Gly1 5 10
15 Glu Thr Val Thr Ile Thr Cys Arg Ala Ser Glu Asn Ile Tyr Asn Asn
20 25 30 Leu Ala Trp Tyr Gln Gln Lys Gln Gly Lys Ser Pro Gln Leu
Leu Val 35 40 45 Tyr Ala Ala Thr Ile Leu Ala Asp Gly Val Pro Ser
Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Gln Tyr Ser Leu Lys
Ile Asn Ser Leu Gln Ser65 70 75 80 Gly Asp Phe Gly Thr Tyr Tyr Cys
Gln His Phe Trp Gly Thr Pro Leu 85 90 95 Thr Phe Gly Ala Gly Thr
Lys Leu Glu Leu Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145
150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser
Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210
131642DNAArtificial SequenceChimeric antibody sequence
131gacatccaga tgacccagag ccccgctagc ctcagcgtgt ccgtcggcga
gaccgtgacc 60atcacctgca gggccagcga gaacatctac aacaacctgg cctggtatca
gcagaagcag 120ggcaaaagcc cccagctgct ggtgtacgcc gccaccattc
tggccgacgg cgtgcccagc 180aggttctctg gaagcggcag cggcacccag
tacagcctga agatcaacag cctgcagagc 240ggggacttcg gcacctacta
ctgccagcac ttctggggca ctcccctgac cttcggagcc 300ggcaccaagc
tggagctgaa gcgtacggtg gccgccccca gcgtgttcat cttccccccc
360agcgatgagc agctgaagag cggcaccgcc agcgtggtgt gtctgctgaa
caacttctac 420ccccgggagg ccaaggtgca gtggaaggtg gacaatgccc
tgcagagcgg caacagccag 480gagagcgtga ccgagcagga cagcaaggac
tccacctaca gcctgagcag caccctgacc 540ctgagcaagg ccgactacga
gaagcacaag gtgtacgcct gtgaggtgac ccaccagggc 600ctgtccagcc
ccgtgaccaa gagcttcaac cggggcgagt gc 642132118PRTmus musculus 132Gln
Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Lys Pro Gly Ala1 5 10
15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45 Gly Ile Ile His Pro Asn Ser Gly Ser Thr Asn Tyr
Asn Glu Lys Phe 50 55 60 Lys Ser Lys Ala Thr Leu Thr Val Asp Lys
Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser
Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Gly Ile Tyr Asp
Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val
Ser Ser 115 133354DNAmus musculus 133caggtgcagc tccagcagcc
cggagccgaa ctggtgaagc ccggagccag cgtcaaactg 60tcctgcaagg ccagcggcta
caccttcacc aactactgga tgcactgggt gaagcagagg 120cccggccagg
gcctggagtg gatcggcatc atccacccca acagcgggag caccaactac
180aacgagaagt tcaagagcaa ggccaccctg accgtggaca agagcagcag
cactgcctac 240atgcagctga gcagcctgac cagcgaggac agcgctgtgt
actactgcgc caggggcatc 300tacgactacc ccttcgccta ttggggccag
ggcacactag tgaccgtgtc cagc 354134448PRTArtificial SequenceChimeric
antibody sequence 134Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu
Val Lys Pro Gly Ala1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser
Gly Tyr Thr Phe Thr Asn Tyr 20 25 30 Trp Met His Trp Val Lys Gln
Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Ile Ile His Pro
Asn Ser Gly Ser Thr Asn Tyr Asn Glu Lys Phe 50 55 60 Lys Ser Lys
Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80 Met
Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Gly Ile Tyr Asp Tyr Pro Phe Ala Tyr Trp Gly Gln Gly Thr
100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val Thr Val Ser Trp Asn145 150 155 160 Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn
Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215
220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
Ser225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr305 310 315 320 Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330
335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu
340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu
Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val Leu Asp385
390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu
Ser Leu Ser Pro Gly Lys 435 440 445 1351344DNAArtificial
SequenceChimeric antibody sequence 135caggtgcagc tccagcagcc
cggagccgaa ctggtgaagc ccggagccag cgtcaaactg 60tcctgcaagg ccagcggcta
caccttcacc aactactgga tgcactgggt gaagcagagg 120cccggccagg
gcctggagtg gatcggcatc atccacccca acagcgggag caccaactac
180aacgagaagt tcaagagcaa ggccaccctg accgtggaca agagcagcag
cactgcctac 240atgcagctga gcagcctgac cagcgaggac agcgctgtgt
actactgcgc caggggcatc 300tacgactacc ccttcgccta ttggggccag
ggcacactag tgaccgtgtc cagcgccagc 360accaagggcc ccagcgtgtt
ccccctggcc cccagcagca agagcaccag cggcggcaca 420gccgccctgg
gctgcctggt gaaggactac ttccccgaac cggtgaccgt gtcctggaac
480agcggagccc tgaccagcgg cgtgcacacc ttccccgccg tgctgcagag
cagcggcctg 540tacagcctga gcagcgtggt gaccgtgccc agcagcagcc
tgggcaccca gacctacatc 600tgtaacgtga accacaagcc cagcaacacc
aaggtggaca agaaggtgga gcccaagagc 660tgtgacaaga cccacacctg
ccccccctgc cctgcccccg agctgctggg aggccccagc 720gtgttcctgt
tcccccccaa gcctaaggac accctgatga tcagcagaac ccccgaggtg
780acctgtgtgg tggtggatgt gagccacgag gaccctgagg tgaagttcaa
ctggtacgtg 840gacggcgtgg aggtgcacaa tgccaagacc aagcccaggg
aggagcagta caacagcacc 900taccgggtgg tgtccgtgct gaccgtgctg
caccaggatt ggctgaacgg caaggagtac 960aagtgtaagg tgtccaacaa
ggccctgcct gcccctatcg agaaaaccat cagcaaggcc 1020aagggccagc
ccagagagcc ccaggtgtac accctgcccc ctagcagaga tgagctgacc
1080aagaaccagg tgtccctgac ctgcctggtg aagggcttct accccagcga
catcgccgtg 1140gagtgggaga gcaacggcca gcccgagaac aactacaaga
ccaccccccc tgtgctggac 1200agcgatggca gcttcttcct gtacagcaag
ctgaccgtgg acaagagcag atggcagcag 1260ggcaacgtgt tcagctgctc
cgtgatgcac gaggccctgc acaatcacta cacccagaag 1320agcctgagcc
tgtcccctgg caag 1344136112PRTmus musculus 136Asp Ile Val Leu Thr
Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1 5 10 15 Gln Arg Ala
Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Ile His 20 25 30 Gly
Thr His Leu Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40
45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Val Pro Ala
50 55 60 Arg Phe Ser Gly Ser Gly Ser Glu Thr Asp Phe Thr Leu Asn
Ile His65 70 75 80 Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Phe Cys
Gln Gln Ser Ile 85 90 95 Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr
Lys Leu Glu Ile Lys Arg 100 105 110 137336DNAmus musculus
137gacatcgtgc tgacccagtc tcccgctagc ctggccgtgt ctctgggcca
gagggccaca 60atcagctgca gggccagcga gagcgtcagc attcacggca cccacctgat
gcactggtac 120cagcagaagc ccggccagcc tcccaagctc ctgatctacg
ccgccagcaa cctggaaagc 180ggagtgcccg ccaggttcag cggcagcggc
tccgagaccg acttcaccct gaacatccac 240cccgtggagg aggaggacgc
cgccacctac ttctgccagc agagcatcga ggacccctac 300accttcggcg
gcggcaccaa gctggagatc aagcgt 336138218PRTArtificial
SequenceChimeric antibody sequence 138Asp Ile Val Leu Thr Gln Ser
Pro Ala Ser Leu Ala Val Ser Leu Gly1 5 10 15 Gln Arg Ala Thr Ile
Ser Cys Arg Ala Ser Glu Ser Val Ser Ile His 20 25 30 Gly Thr His
Leu Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 35 40 45 Lys
Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser Gly Val Pro Ala 50 55
60 Arg Phe Ser Gly Ser Gly Ser Glu Thr Asp Phe Thr Leu Asn Ile
His65 70 75 80 Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Phe Cys Gln
Gln Ser Ile 85 90 95 Glu Asp Pro Tyr Thr Phe Gly Gly Gly Thr Lys
Leu Glu Ile Lys Arg 100 105 110 Thr 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 Ser145 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
139654DNAArtificial SequenceChimeric antibody sequence
139gacatcgtgc tgacccagtc tcccgctagc ctggccgtgt ctctgggcca
gagggccaca 60atcagctgca gggccagcga gagcgtcagc attcacggca cccacctgat
gcactggtac 120cagcagaagc ccggccagcc tcccaagctc ctgatctacg
ccgccagcaa cctggaaagc 180ggagtgcccg ccaggttcag cggcagcggc
tccgagaccg acttcaccct gaacatccac 240cccgtggagg aggaggacgc
cgccacctac ttctgccagc agagcatcga ggacccctac 300accttcggcg
gcggcaccaa gctggagatc aagcgtacgg tggccgcccc cagcgtgttc
360atcttccccc ccagcgatga gcagctgaag agcggcaccg ccagcgtggt
gtgtctgctg 420aacaacttct acccccggga ggccaaggtg cagtggaagg
tggacaatgc cctgcagagc 480ggcaacagcc aggagagcgt gaccgagcag
gacagcaagg actccaccta cagcctgagc 540agcaccctga ccctgagcaa
ggccgactac gagaagcaca aggtgtacgc ctgtgaggtg 600acccaccagg
gcctgtccag ccccgtgacc aagagcttca accggggcga gtgc 654140120PRTmus
musculus 140Glu Val Lys Leu Leu Gln Ser Gly Gly Gly Leu Val Gln Pro
Gly Gly1 5 10 15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Ile Asp
Phe Ser Arg Tyr 20 25 30 Trp Met Ser Trp Val Arg Arg Ala Pro Gly
Lys Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Asp Arg Ser
Thr Ile Asn Tyr Ala Pro Ser Leu 50 55 60 Lys Asp Lys Phe Ile Ile
Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr65 70 75 80 Leu Gln Met Ser
Lys Val Arg Ser Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Val
Phe Tyr Tyr Asp Tyr Glu Gly Ala Met Asp Tyr Trp Gly Gln 100 105 110
Gly Thr Ser Val Thr Val Ser Ser 115 120 141360DNAmus musculus
141gaggtgaagc ttctccagtc tggaggtggc ctggtgcagc ctggaggatc
cctgaaactc 60tcctgtgcag cctcaggaat cgattttagt agatactgga tgagttgggt
tcggcgggct 120ccagggaaag gactagaatg gattggagaa attaatccag
ataggagtac aatcaactat 180gcaccatctc taaaggataa attcatcatc
tccagagaca acgccaaaaa tacgctgtac 240ctgcaaatga gcaaagtgag
atctgaggac acagcccttt attactgtgc agttttctac 300tatgattacg
agggtgctat ggactactgg ggtcaaggaa cctcagtcac cgtctcctca
360142450PRTArtificial SequenceChimeric antibody sequence 142Glu
Val Lys Leu Leu Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10
15 Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Ile Asp Phe Ser Arg Tyr
20 25 30 Trp Met Ser Trp Val Arg Arg Ala Pro Gly Lys Gly Leu Glu
Trp Ile 35 40 45 Gly Glu Ile Asn Pro Asp Arg Ser Thr Ile Asn Tyr
Ala Pro Ser Leu 50 55 60 Lys Asp Lys Phe Ile Ile Ser Arg Asp Asn
Ala Lys Asn Thr Leu Tyr65 70 75 80 Leu Gln Met Ser Lys Val Arg Ser
Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Val Phe Tyr Tyr Asp
Tyr Glu Gly Ala Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Ser Val
Thr Val Ser Ser Ala Lys Thr Thr Ala Pro Ser Val 115 120 125 Phe Pro
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser145
150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile
Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys
Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro
Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly225 230 235 240 Pro Ser Val
Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser
Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265
270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr
Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp
Leu Asn Gly Lys305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu Pro Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser
Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu
Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390
395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 1431350DNAArtificial
SequenceChimeric antibody sequence 143gaggtgaagc ttctccagtc
tggaggtggc ctggtgcagc ctggaggatc cctgaaactc 60tcctgtgcag cctcaggaat
cgattttagt agatactgga tgagttgggt tcggcgggct 120ccagggaaag
gactagaatg gattggagaa attaatccag ataggagtac aatcaactat
180gcaccatctc taaaggataa attcatcatc tccagagaca acgccaaaaa
tacgctgtac 240ctgcaaatga gcaaagtgag atctgaggac acagcccttt
attactgtgc agttttctac 300tatgattacg agggtgctat ggactactgg
ggtcaaggaa cctcagtcac cgtctcctca 360gccaaaacaa cagcccccag
cgtgttcccc ctggccccca gcagcaagag caccagcggc 420ggcacagccg
ccctgggctg cctggtgaag gactacttcc ccgaaccggt gaccgtgtcc
480tggaacagcg gagccctgac cagcggcgtg cacaccttcc ccgccgtgct
gcagagcagc 540ggcctgtaca gcctgagcag cgtggtgacc gtgcccagca
gcagcctggg cacccagacc 600tacatctgta acgtgaacca caagcccagc
aacaccaagg tggacaagaa ggtggagccc 660aagagctgtg acaagaccca
cacctgcccc ccctgccctg cccccgagct gctgggaggc 720cccagcgtgt
tcctgttccc ccccaagcct aaggacaccc tgatgatcag cagaaccccc
780gaggtgacct gtgtggtggt ggatgtgagc cacgaggacc ctgaggtgaa
gttcaactgg 840tacgtggacg gcgtggaggt gcacaatgcc aagaccaagc
ccagggagga gcagtacaac 900agcacctacc gggtggtgtc cgtgctgacc
gtgctgcacc aggattggct gaacggcaag 960gagtacaagt gtaaggtgtc
caacaaggcc ctgcctgccc ctatcgagaa aaccatcagc 1020aaggccaagg
gccagcccag agagccccag gtgtacaccc tgccccctag cagagatgag
1080ctgaccaaga accaggtgtc cctgacctgc ctggtgaagg gcttctaccc
cagcgacatc 1140gccgtggagt gggagagcaa cggccagccc gagaacaact
acaagaccac cccccctgtg 1200ctggacagcg atggcagctt cttcctgtac
agcaagctga ccgtggacaa gagcagatgg 1260cagcagggca acgtgttcag
ctgctccgtg atgcacgagg ccctgcacaa tcactacacc 1320cagaagagcc
tgagcctgtc ccctggcaag 1350144108PRTmus musculus 144Asp Ile Val Met
Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10 15 Asp Arg
Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Asp Thr Asn 20 25 30
Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala Leu Ile 35
40 45 Tyr Ser Ala Ser Tyr Arg Phe Ser Gly Val Pro Asp Arg Phe Thr
Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Asn
Val Gln Ser65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys Gln Gln Tyr
Asn Ser Phe Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr Lys Leu Glu
Ile Lys Arg 100 105 145324DNAmus musculus 145gacattgtga tgacccagtc
tcaaaaattc atgtccacat cagtaggaga cagggtcagc 60gtcacctgca aggccagtca
gaatgtggat actaatgtag cctggtatca acaaaaacca 120gggcaatctc
ctaaagcact gatttactcg gcatcctacc ggttcagtgg agtccctgat
180cgcttcacag gcagtggatc tgggacagat ttcactctca ccatcagcaa
tgtgcagtct 240gaagacttgg cagagtattt ctgtcagcaa tataacagct
ttccattcac gttcggctcg 300gggacaaagt tggaaataaa acgt
324146214PRTArtificial SequenceChimeric antibody sequence 146Asp
Ile Val Met Thr Gln Ser Gln Lys Phe Met Ser Thr Ser Val Gly1 5 10
15 Asp Arg Val Ser Val Thr Cys Lys Ala Ser Gln Asn Val Asp Thr Asn
20 25 30 Val Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ser Pro Lys Ala
Leu Ile 35 40 45 Tyr Ser Ala Ser Tyr Arg Phe Ser Gly Val Pro Asp
Arg Phe Thr Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser Asn Val Gln Ser65 70 75 80 Glu Asp Leu Ala Glu Tyr Phe Cys
Gln Gln Tyr Asn Ser Phe Pro Phe 85 90 95 Thr Phe Gly Ser Gly Thr
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140
Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145
150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser
Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys
His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser
Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210
147642PRTArtificial SequenceChimeric antibody sequence 147Gly Ala
Cys Ala Thr Thr Gly Thr Gly Ala Thr Gly Ala Cys Cys Cys1 5 10 15
Ala Gly Thr Cys Thr Cys Ala Ala Ala Ala Ala Thr Thr Cys Ala Thr 20
25 30 Gly Thr Cys Cys Ala Cys Ala Thr Cys Ala Gly Thr Ala Gly Gly
Ala 35 40 45 Gly Ala Cys Ala Gly Gly Gly Thr Cys Ala Gly Cys Gly
Thr Cys Ala 50 55 60 Cys Cys Thr Gly Cys Ala Ala Gly Gly Cys Cys
Ala Gly Thr Cys Ala65 70 75 80 Gly Ala Ala Thr Gly Thr Gly Gly Ala
Thr Ala Cys Thr Ala Ala Thr 85 90 95 Gly Thr Ala Gly Cys Cys Thr
Gly Gly Thr Ala Thr Cys Ala Ala Cys 100 105 110 Ala Ala Ala Ala Ala
Cys Cys Ala Gly Gly Gly Cys Ala Ala Thr Cys 115 120 125 Thr Cys Cys
Thr Ala Ala Ala Gly Cys Ala Cys Thr Gly Ala Thr Thr 130 135 140 Thr
Ala Cys Thr Cys Gly Gly Cys Ala Thr Cys Cys Thr Ala Cys Cys145 150
155 160 Gly Gly Thr Thr Cys Ala Gly Thr Gly Gly Ala Gly Thr Cys Cys
Cys 165 170 175 Thr Gly Ala Thr Cys Gly Cys Thr Thr Cys Ala Cys Ala
Gly Gly Cys 180 185 190 Ala Gly Thr Gly Gly Ala Thr Cys Thr Gly Gly
Gly Ala Cys Ala Gly 195 200 205 Ala Thr Thr Thr Cys Ala Cys Thr Cys
Thr Cys Ala Cys Cys Ala Thr 210 215 220 Cys Ala Gly Cys Ala Ala Thr
Gly Thr Gly Cys Ala Gly Thr Cys Thr225 230 235 240 Gly Ala Ala Gly
Ala Cys Thr Thr Gly Gly Cys Ala Gly Ala Gly Thr 245 250 255 Ala Thr
Thr Thr Cys Thr Gly Thr Cys Ala Gly Cys Ala Ala Thr Ala 260 265
270 Thr Ala Ala Cys Ala Gly Cys Thr Thr Thr Cys Cys Ala Thr Thr Cys
275 280 285 Ala Cys Gly Thr Thr Cys Gly Gly Cys Thr Cys Gly Gly Gly
Gly Ala 290 295 300 Cys Ala Ala Ala Gly Thr Thr Gly Gly Ala Ala Ala
Thr Ala Ala Ala305 310 315 320 Ala Cys Gly Thr Ala Cys Gly Gly Thr
Gly Gly Cys Cys Gly Cys Cys 325 330 335 Cys Cys Cys Ala Gly Cys Gly
Thr Gly Thr Thr Cys Ala Thr Cys Thr 340 345 350 Thr Cys Cys Cys Cys
Cys Cys Cys Ala Gly Cys Gly Ala Thr Gly Ala 355 360 365 Gly Cys Ala
Gly Cys Thr Gly Ala Ala Gly Ala Gly Cys Gly Gly Cys 370 375 380 Ala
Cys Cys Gly Cys Cys Ala Gly Cys Gly Thr Gly Gly Thr Gly Thr385 390
395 400 Gly Thr Cys Thr Gly Cys Thr Gly Ala Ala Cys Ala Ala Cys Thr
Thr 405 410 415 Cys Thr Ala Cys Cys Cys Cys Cys Gly Gly Gly Ala Gly
Gly Cys Cys 420 425 430 Ala Ala Gly Gly Thr Gly Cys Ala Gly Thr Gly
Gly Ala Ala Gly Gly 435 440 445 Thr Gly Gly Ala Cys Ala Ala Thr Gly
Cys Cys Cys Thr Gly Cys Ala 450 455 460 Gly Ala Gly Cys Gly Gly Cys
Ala Ala Cys Ala Gly Cys Cys Ala Gly465 470 475 480 Gly Ala Gly Ala
Gly Cys Gly Thr Gly Ala Cys Cys Gly Ala Gly Cys 485 490 495 Ala Gly
Gly Ala Cys Ala Gly Cys Ala Ala Gly Gly Ala Cys Thr Cys 500 505 510
Cys Ala Cys Cys Thr Ala Cys Ala Gly Cys Cys Thr Gly Ala Gly Cys 515
520 525 Ala Gly Cys Ala Cys Cys Cys Thr Gly Ala Cys Cys Cys Thr Gly
Ala 530 535 540 Gly Cys Ala Ala Gly Gly Cys Cys Gly Ala Cys Thr Ala
Cys Gly Ala545 550 555 560 Gly Ala Ala Gly Cys Ala Cys Ala Ala Gly
Gly Thr Gly Thr Ala Cys 565 570 575 Gly Cys Cys Thr Gly Thr Gly Ala
Gly Gly Thr Gly Ala Cys Cys Cys 580 585 590 Ala Cys Cys Ala Gly Gly
Gly Cys Cys Thr Gly Thr Cys Cys Ala Gly 595 600 605 Cys Cys Cys Cys
Gly Thr Gly Ala Cys Cys Ala Ala Gly Ala Gly Cys 610 615 620 Thr Thr
Cys Ala Ala Cys Cys Gly Gly Gly Gly Cys Gly Ala Gly Thr625 630 635
640 Gly Cys148118PRTmus musculus 148Pro Val Gln Leu Gln Gln Pro Gly
Thr Glu Leu Val Arg Pro Gly Thr1 5 10 15 Ser Val Lys Leu Ser Cys
Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Trp Met His Trp
Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val
Ile Asp Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60
Lys Gly Lys Ala Thr Leu Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr65
70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Gln Val Phe Asp Tyr Pro Met Asp Tyr Trp
Gly Gln Gly Thr 100 105 110 Ser Val Thr Val Ser Ser 115
149354DNAmus musculus 149ccggtccaac tgcagcagcc tgggactgag
ctggtgaggc ctgggacttc agtgaagttg 60tcctgcaagg cttctggcta caccttcacc
agctactgga tgcactgggt aaagcagagg 120cctggacaag gccttgagtg
gatcggagtg attgatcctt ctgatagtta tactaactac 180aatcaaaagt
tcaagggcaa ggccacattg actgtagaca catcctccag cacagcctac
240atgcagctca gcagcctgac atctgaggac tctgcggtct attactgtgc
aagacaggtg 300tttgactatc ctatggacta ctggggtcaa ggaacctcag
tcaccgtctc ctca 354150448PRTArtificial SequenceChimeric antibody
sequence 150Pro Val Gln Leu Gln Gln Pro Gly Thr Glu Leu Val Arg Pro
Gly Thr1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Ser His 20 25 30 Trp Met His Trp Val Lys Gln Arg Pro Gly
Gln Gly Leu Glu Trp Ile 35 40 45 Gly Val Ile Asp Pro Ser Asp Ser
Tyr Thr Asn Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Thr Leu
Thr Val Asp Thr Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Gln Leu Ser
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg
Gln Val Phe Asp Tyr Pro Met Asp Tyr Trp Gly Gln Gly Thr 100 105 110
Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115
120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val
Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser225 230 235
240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr305 310 315 320 Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro
Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360
365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
1511344DNAArtificial SequenceChimeric antibody sequence
151ccggtccaac tgcagcagcc tgggactgag ctggtgaggc ctgggacttc
agtgaagttg 60tcctgcaagg cttctggcta caccttcacc agccactgga tgcactgggt
aaagcagagg 120cctggacaag gccttgagtg gatcggagtg attgatcctt
ctgatagtta tactaactac 180aatcaaaagt tcaagggcaa ggccacattg
actgtagaca catcctccag cacagcctac 240atgcagctca gcagcctgac
atctgaggac tctgcggtct attactgtgc aagacaggtg 300tttgactatc
ctatggacta ctggggtcaa ggaacactag tgaccgtgtc cagcgccagc
360accaagggcc ccagcgtgtt ccccctggcc cccagcagca agagcaccag
cggcggcaca 420gccgccctgg gctgcctggt gaaggactac ttccccgaac
cggtgaccgt gtcctggaac 480agcggagccc tgaccagcgg cgtgcacacc
ttccccgccg tgctgcagag cagcggcctg 540tacagcctga gcagcgtggt
gaccgtgccc agcagcagcc tgggcaccca gacctacatc 600tgtaacgtga
accacaagcc cagcaacacc aaggtggaca agaaggtgga gcccaagagc
660tgtgacaaga cccacacctg ccccccctgc cctgcccccg agctgctggg
aggccccagc 720gtgttcctgt tcccccccaa gcctaaggac accctgatga
tcagcagaac ccccgaggtg 780acctgtgtgg tggtggatgt gagccacgag
gaccctgagg tgaagttcaa ctggtacgtg 840gacggcgtgg aggtgcacaa
tgccaagacc aagcccaggg aggagcagta caacagcacc 900taccgggtgg
tgtccgtgct gaccgtgctg caccaggatt ggctgaacgg caaggagtac
960aagtgtaagg tgtccaacaa ggccctgcct gcccctatcg agaaaaccat
cagcaaggcc 1020aagggccagc ccagagagcc ccaggtgtac accctgcccc
ctagcagaga tgagctgacc 1080aagaaccagg tgtccctgac ctgcctggtg
aagggcttct accccagcga catcgccgtg 1140gagtgggaga gcaacggcca
gcccgagaac aactacaaga ccaccccccc tgtgctggac 1200agcgatggca
gcttcttcct gtacagcaag ctgaccgtgg acaagagcag atggcagcag
1260ggcaacgtgt tcagctgctc cgtgatgcac gaggccctgc acaatcacta
cacccagaag 1320agcctgagcc tgtcccctgg caag 1344152112PRTmus musculus
152Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1
5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Ile
His 20 25 30 Gly Thr His Leu Met His Trp Tyr Gln Gln Lys Pro Gly
Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu
Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Glu Thr
Asp Phe Thr Leu Asn Ile His65 70 75 80 Pro Val Glu Glu Glu Asp Ala
Ala Thr Tyr Phe Cys Gln Gln Ser Ile 85 90 95 Glu Asp Pro Trp Thr
Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg 100 105 110
153336DNAmus musculus 153gacattgtgc tgacccaatc tccagcttct
ttggctgtgt ctctagggca gagggccacc 60atctcctgca gagccagtga aagtgtcagt
attcatggta ctcatttaat gcactggtac 120caacagaaac caggacagcc
acccaaactc ctcatctatg ctgcatccaa cctagaatct 180ggagtccctg
ccaggttcag tggcagtggg tctgagacag acttcaccct caacatccat
240cctgtggagg aggaggatgc tgcaacctat ttctgtcagc aaagtattga
ggatccgtgg 300acgttcggtg gaggcaccaa gctggaaatc aaacgt
336154218PRTArtificial SequenceChimeric antibody sequence 154Asp
Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1 5 10
15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Ile His
20 25 30 Gly Thr His Leu Met His Trp Tyr Gln Gln Lys Pro Gly Gln
Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser
Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Ser Gly Ser Glu Thr Asp
Phe Thr Leu Asn Ile His65 70 75 80 Pro Val Glu Glu Glu Asp Ala Ala
Thr Tyr Phe Cys Gln Gln Ser Ile 85 90 95 Glu Asp Pro Trp Thr Phe
Gly Gly Gly Thr Lys Leu Glu Ile Asn Arg 100 105 110 Thr 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 Ser145
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 155654DNAArtificial SequenceChimeric antibody
sequence 155gacattgtgc tgacccaatc tccagcttct ttggctgtgt ctctagggca
gagggccacc 60atctcctgca gagccagtga aagtgtcagt attcatggta ctcatttaat
gcactggtac 120caacagaaac caggacagcc acccaaactc ctcatctatg
ctgcatccaa cctagaatct 180ggagtccctg ccaggttcag tggcagtggg
tctgagacag acttcaccct caacatccat 240cctgtggagg aggaggatgc
tgcaacctat ttctgtcagc aaagtattga ggatccgtgg 300acgttcggtg
gaggcaccaa gctggaaatc aatcgtacgg tggccgcccc cagcgtgttc
360atcttccccc ccagcgatga gcagctgaag agcggcaccg ccagcgtggt
gtgtctgctg 420aacaacttct acccccggga ggccaaggtg cagtggaagg
tggacaatgc cctgcagagc 480ggcaacagcc aggagagcgt gaccgagcag
gacagcaagg actccaccta cagcctgagc 540agcaccctga ccctgagcaa
ggccgactac gagaagcaca aggtgtacgc ctgtgaggtg 600acccaccagg
gcctgtccag ccccgtgacc aagagcttca accggggcga gtgc 654156118PRTmus
musculus 156Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro
Gly Ala1 5 10 15 Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Asp Tyr 20 25 30 Glu Met His Trp Val Lys Gln Thr Pro Val
His Gly Leu Glu Trp Ile 35 40 45 Gly Ala Ile Asp Pro Glu Thr Gly
Gly Thr Ala Tyr Asn Gln Lys Phe 50 55 60 Lys Gly Lys Ala Ile Leu
Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr65 70 75 80 Met Glu Leu Arg
Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Thr Arg
Ser Ile Tyr Asp Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110
Thr Leu Thr Val Ser Ser 115 157354DNAmus musculus 157caggttcaac
tgcagcagtc tggggctgag ctggtgaggc ctggggcttc agtgacgctg 60tcctgcaagg
cttcgggcta cacatttact gactatgaaa tgcactgggt gaagcagaca
120cctgtgcatg gcctggaatg gattggagct attgatcctg aaactggtgg
tactgcctac 180aatcagaagt tcaagggcaa ggccatactg actgcagaca
aatcctccag cacagcctac 240atggagctcc gcagcctgac atctgaggac
tctgccgtct attactgtac aagatcgatt 300tatgattact actttgacta
ctggggccaa ggcaccactc tcacagtctc ctca 354158448PRTArtificial
SequenceChimeric antibody sequence 158Gln Val Gln Leu Gln Gln Ser
Gly Ala Glu Leu Val Arg Pro Gly Ala1 5 10 15 Ser Val Thr Leu Ser
Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 20 25 30 Glu Met His
Trp Val Lys Gln Thr Pro Val His Gly Leu Glu Trp Ile 35 40 45 Gly
Ala Ile Asp Pro Glu Thr Gly Gly Thr Ala Tyr Asn Gln Lys Phe 50 55
60 Lys Gly Lys Ala Ile Leu Thr Ala Asp Lys Ser Ser Ser Thr Ala
Tyr65 70 75 80 Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val
Tyr Tyr Cys 85 90 95 Thr Arg Ser Ile Tyr Asp Tyr Tyr Phe Asp Tyr
Trp Gly Gln Gly Thr 100 105 110 Thr Leu Thr Val Ser Ser Ala Lys Thr
Thr Pro Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser
Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn145 150 155 160 Ser Gly
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180
185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys
Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
Leu Gly Gly Pro Ser225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr305
310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn
Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser
Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn
Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445
1591344DNAArtificial SequenceChimeric antibody sequence
159caggttcaac tgcagcagtc tggggctgag ctggtgaggc ctggggcttc
agtgacgctg 60tcctgcaagg cttcgggcta cacatttact gactatgaaa tgcactgggt
gaagcagaca 120cctgtgcatg gcctggaatg gattggagct attgatcctg
aaactggtgg tactgcctac 180aatcagaagt tcaagggcaa ggccatactg
actgcagaca aatcctccag cacagcctac 240atggagctcc gcagcctgac
atctgaggac tctgccgtct attactgtac aagatcgatt 300tatgattact
actttgacta ctggggccaa ggcaccactc tcacagtctc ctcagccaaa
360acgacacccc ccagcgtgtt ccccctggcc cccagcagca agagcaccag
cggcggcaca 420gccgccctgg gctgcctggt gaaggactac ttccccgaac
cggtgaccgt gtcctggaac 480agcggagccc tgaccagcgg cgtgcacacc
ttccccgccg tgctgcagag cagcggcctg 540tacagcctga gcagcgtggt
gaccgtgccc agcagcagcc tgggcaccca gacctacatc 600tgtaacgtga
accacaagcc cagcaacacc aaggtggaca agaaggtgga gcccaagagc
660tgtgacaaga cccacacctg ccccccctgc cctgcccccg agctgctggg
aggccccagc 720gtgttcctgt tcccccccaa gcctaaggac accctgatga
tcagcagaac ccccgaggtg 780acctgtgtgg tggtggatgt gagccacgag
gaccctgagg tgaagttcaa ctggtacgtg 840gacggcgtgg aggtgcacaa
tgccaagacc aagcccaggg aggagcagta caacagcacc 900taccgggtgg
tgtccgtgct gaccgtgctg caccaggatt ggctgaacgg caaggagtac
960aagtgtaagg tgtccaacaa ggccctgcct gcccctatcg agaaaaccat
cagcaaggcc 1020aagggccagc ccagagagcc ccaggtgtac accctgcccc
ctagcagaga tgagctgacc 1080aagaaccagg tgtccctgac ctgcctggtg
aagggcttct accccagcga catcgccgtg 1140gagtgggaga gcaacggcca
gcccgagaac aactacaaga ccaccccccc tgtgctggac 1200agcgatggca
gcttcttcct gtacagcaag ctgaccgtgg acaagagcag atggcagcag
1260ggcaacgtgt tcagctgctc cgtgatgcac gaggccctgc acaatcacta
cacccagaag 1320agcctgagcc tgtcccctgg caag 1344160112PRTmus musculus
160Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1
5 10 15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Ile
His 20 25 30 Gly Thr His Leu Met His Trp Tyr Gln Gln Lys Pro Gly
Gln Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu
Ser Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Gly Gly Ser Glu Thr
Asp Phe Thr Leu Asn Ile His65 70 75 80 Pro Val Glu Glu Glu Asp Gly
Ala Thr Tyr Phe Cys Gln Gln Ser Ile 85 90 95 Glu Tyr Pro Arg Thr
Phe Gly Gly Gly Thr Lys Leu Glu Ile Asn Arg 100 105 110
161336DNAmus musculus 161gacattgtgc tgacccaatc tccagcttct
ttggctgtgt ctctagggca gagggccacc 60atctcctgca gagccagtga aagtgtcagt
attcatggta ctcatttaat gcactggtac 120caacagaaac caggacagcc
acccaaactc ctcatctatg ctgcatccaa cctagaatct 180ggagtccctg
ccaggttcag tggcggtggg tctgagacag acttcaccct caacatccat
240cctgtggagg aggaggatgg tgcaacctat ttctgtcagc aaagtattga
gtatcctcgg 300acgttcggtg gaggcaccaa gctggaaatc aatcgt
336162218PRTArtificial SequenceChimeric antibody sequence 162Asp
Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly1 5 10
15 Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Glu Ser Val Ser Ile His
20 25 30 Gly Thr His Leu Met His Trp Tyr Gln Gln Lys Pro Gly Gln
Pro Pro 35 40 45 Lys Leu Leu Ile Tyr Ala Ala Ser Asn Leu Glu Ser
Gly Val Pro Ala 50 55 60 Arg Phe Ser Gly Gly Gly Ser Glu Thr Asp
Phe Thr Leu Asn Ile His65 70 75 80 Pro Val Glu Glu Glu Asp Gly Ala
Thr Tyr Phe Cys Gln Gln Ser Ile 85 90 95 Glu Tyr Pro Arg Thr Phe
Gly Gly Gly Thr Lys Leu Glu Ile Asn Arg 100 105 110 Thr 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 Ser145
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 163654DNAArtificial SequenceChimeric antibody
sequence 163gacattgtgc tgacccaatc tccagcttct ttggctgtgt ctctagggca
gagggccacc 60atctcctgca gagccagtga aagtgtcagt attcatggta ctcatttaat
gcactggtac 120caacagaaac caggacagcc acccaaactc ctcatctatg
ctgcatccaa cctagaatct 180ggagtccctg ccaggttcag tggcggtggg
tctgagacag acttcaccct caacatccat 240cctgtggagg aggaggatgg
tgcaacctat ttctgtcagc aaagtattga gtatcctcgg 300acgttcggtg
gaggcaccaa gctggaaatc aatcgtacgg tggccgcccc cagcgtgttc
360atcttccccc ccagcgatga gcagctgaag agcggcaccg ccagcgtggt
gtgtctgctg 420aacaacttct acccccggga ggccaaggtg cagtggaagg
tggacaatgc cctgcagagc 480ggcaacagcc aggagagcgt gaccgagcag
gacagcaagg actccaccta cagcctgagc 540agcaccctga ccctgagcaa
ggccgactac gagaagcaca aggtgtacgc ctgtgaggtg 600acccaccagg
gcctgtccag ccccgtgacc aagagcttca accggggcga gtgc 6541645PRTmus
musculus 164Asp Tyr Tyr Asn Met1 5 16516PRTmus musculus 165Val Ile
Asn Pro Tyr Asn Gly Gly Thr Asp Tyr Asn Gln Lys Phe Gly1 5 10 15
1669PRTmus musculus 166Ser Val Tyr Asp Tyr Pro Phe Asp Tyr1 5
16715PRTmus musculus 167Arg Ala Ser Glu Ser Val Ser Ile His Gly Thr
His Leu Met His1 5 10 15 1687PRTmus musculus 168Ala Ala Ser Asn Leu
Glu Ser1 5 1699PRTmus musculus 169Gln Gln Ser Ile Glu Asp Pro Arg
Thr1 5 1705PRTmus musculus 170Asp Tyr Ser Ile Asp1 5 17117PRTmus
musculus 171Asp Ile Asp Pro Asn Tyr Gly Asp Pro Ile Tyr Asn His Lys
Phe Lys1 5 10 15 Gly17210PRTmus musculus 172Arg Ala Thr Gly Thr Asp
Trp Phe Ala Phe1 5 10 17311PRTmus musculus 173Arg Ala Ser Glu Asn
Ile Tyr Asn Asn Leu Ala1 5 10 1747PRTmus musculus 174Ala Ala Thr
Ile Leu Ala Asp1 5 1759PRTmus musculus 175Gln His Phe Trp Gly Thr
Pro Leu Thr1 5 1765PRTmus musculus 176Asn Tyr Trp Met His1 5
17717PRTmus musculus 177Ile Ile His Pro Asn Ser Gly Ser Thr Asn Tyr
Asn Glu Lys Phe Lys1 5 10 15 Ser1789PRTmus musculus 178Gly Ile Tyr
Asp Tyr Pro Phe Ala Tyr1 5 17915PRTmus musculus 179Arg Ala Ser Glu
Ser Val Ser Ile His Gly Thr His Leu Met His1 5 10 15 1807PRTmus
musculus 180Ala Ala Ser Asn Leu Glu Ser1 5 1819PRTmus musculus
181Gln Gln Ser Ile Glu Asp Pro Tyr Thr1 5 1825PRTmus musculus
182Arg Tyr Trp Met Ser1 5 18317PRTmus musculus 183Glu Ile Asn Pro
Asp Arg Ser Thr Ile Asn Tyr Ala Pro Ser Leu Lys1 5 10 15
Asp18411PRTmus musculus 184Phe Tyr Tyr Asp Tyr Glu Gly Ala Met Asp
Tyr1 5 10 18511PRTmus musculus 185Lys Ala Ser Gln Asn Val Asp Thr
Asn Val Ala1 5 10 1867PRTmus musculus 186Ser Ala Ser Tyr Arg Phe
Ser1 5 1879PRTmus musculus 187Gln Gln Tyr Asn Ser Phe Pro Phe Thr1
5 1885PRTmus musculus 188Ser Tyr Trp Met His1 5 18917PRTmus
musculus 189Val Ile Asp Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys
Phe Lys1 5 10 15 Gly1909PRTmus musculus 190Gln Val Phe Asp Tyr Pro
Met Asp Tyr1 5 19115PRTmus musculus 191Arg Ala Ser Glu Ser Val Ser
Ile His Gly Thr His Leu Met His1 5 10 15 1927PRTmus musculus 192Ala
Ala Ser Asn Leu Glu Ser1 5 1939PRTmus musculus 193Gln Gln Ser Ile
Glu Asp Pro Trp Thr1 5 1945PRTmus musculus 194Asp Tyr Glu Met His1
5 19517PRTmus musculus 195Ala Ile Asp Pro Glu Thr Gly Gly Thr Ala
Tyr Asn Gln Lys Phe Lys1 5 10 15 Gly1969PRTmus musculus 196Ser Ile
Tyr Asp Tyr Tyr Phe Asp Tyr1 5 19715PRTmus musculus 197Arg Ala Ser
Glu Ser Val Ser Ile His Gly Thr His Leu Met His1 5 10 15 1987PRTmus
musculus 198Ala Ala Ser Asn Leu Glu Ser1 5 1999PRTmus musculus
199Gln Gln Ser Ile Glu Tyr Pro Arg Thr1 5
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