Biepitope tetravalent antibody targeting EGFR

Abstract

A biepitope tetravalent antibody having a binding affinity for at least two epitopes of EGFR. The antibody comprises an antibody skeleton and an scFv structural domain connected with the antibody skeleton. The two epitopes of the EGFR may include an EGFR wild type (EGFRwt) epitope and an EGFRvIII epitope. The antibody has higher binding affinity to an EGFRvIII epitope than to an EGFRwt epitope.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims the benefit of the filing date of U.S. Provisional Application Serial No. 63 / 402,945, filed on August 31, 2022, under 35 U.S.C. 119(e), the entire disclosure of which is incorporated herein by reference. Technical Field

[0003] The present disclosure generally relates to the technical field of antibody cancer therapies, and more particularly to bis - epitope tetravalent antibodies. Background Art

[0004] Glioblastoma multiforme is an aggressive cancer that accounts for the majority of malignant tumors originating in the brain and has a poor prognosis, with an average survival time of 14 - 15 months after diagnosis. 1 Most glioblastoma cases (34 - 63%) involve amplification of the EGFR gene. 2 In addition to overexpression of the wild - type EGFR protein, mutant forms of EGFR occur in 63 - 75% of cases, with the most common mutation, known as EGFRvIII, occurring in 25 - 64% of cases. 2 Structurally, the EGFRvIII mutation results in the deletion of 267 residues in the extracellular domain of the protein, which may functionally induce constitutive signaling due to the lower efficiency of receptor endocytosis. 2 These changes cause tumors carrying EGFRvIII to exhibit increased proliferation, increased angiogenesis, and decreased apoptosis. 2 In addition to its association with glioblastoma multiforme, wild - type EGFR and mutant EGFRvIII can also be overexpressed in other types of solid tumors, including ovarian cancer, breast cancer, and lung cancer. 2

[0005] Several EGFR - targeted antibodies, including Cetuximab, Panitumumab, and Necitumumab, have been approved by the FDA for the treatment of epithelial tumors. Nimotuzumab has also been approved in several countries for the treatment of solid tumors. The lower affinity of Nimotuzumab for EGFR compared to Cetuximab or Panitumumab may help explain its reduced skin toxicity. 3Meanwhile, antibody-based therapies specifically targeting mutant EGFRvIII have been evaluated in clinical trials, including the monoclonal antibody Depatuxizumab (Phase I) and the antibody-drug conjugate Depatuxizumab mafodotin (Phase 3). In particular, the Phase 3 trial of the EGFRvIII-targeted ADC was stopped after failing to reach the primary endpoint of overall survival in patients with newly diagnosed glioblastoma. 4 T-cell conjugates targeting EGFRvIII and CD3 have also been preclinically evaluated, further highlighting the promise of EGFRvIII as a therapeutic target. 5,6 Although EGFRvIII-targeted therapies will not be available soon, there is currently no single therapy that simultaneously targets tumor cells expressing oncogenic EGFR, including overexpressed (wt) and mutant EGFR (vIII).

[0006] Potential drawbacks of cetuximab-derived antibodies and ABT806-derived antibodies are that the variable regions of these antibodies are found in mice and retain non-human sequences. It has been demonstrated that chimeric antibodies may have increased immunogenic capabilities when compared to humanized or human antibodies. 7 On the other hand, humanization can not only reduce immunogenicity but also improve the stability of the antibody by making the variable and constant regions more compatible. 8

[0007] The high affinity of cetuximab for its antigen EGFR may lead to off-tumor binding to the target. As a result, toxicities such as severe skin irritation are often observed. Meanwhile, nimotuzumab, which binds to EGFR with lower affinity, seems to have fewer of these undesirable side effects. 9 Therefore, it may be advantageous to reduce the affinity of an antibody such as cetuximab in order to reduce binding to healthy tissues with low expression while retaining its antigen specificity, thereby improving the therapeutic index. In the case of bispecific antibodies, reduced affinity can be used to balance the relative strength of the two binding domains and effectively optimize the overall specificity of the antibody. SUMMARY OF THE INVENTION

[0008] The following summary of the invention is illustrative only and is not intended to be limiting in any way. Other aspects, embodiments, and features will become apparent from the following detailed description when taken in conjunction with the accompanying drawings, in addition to the above-described illustrative aspects, embodiments, and features.

[0009] The present disclosure provides bispecific tetravalent antibodies that target at least two epitopes of EGFR, as well as methods for preparing and using the antibodies.

[0010] In one aspect, the present application provides a bispecific tetravalent antibody having binding affinity for at least two epitopes of EGFR. The antibody may comprise an antibody backbone comprising an antibody light chain having an antibody light chain variable (VL) domain and an antibody heavy chain having an antibody heavy chain variable (VH) domain, wherein the antibody VL domain and the antibody VH domain form a Fab region. The antibody may further comprise an scFv domain having an scFv light chain variable (VL) domain and an scFv heavy chain variable (VH) domain, wherein the scFv domain is linked to at least one end of the antibody light chain or the antibody heavy chain via a domain linker.

[0011] In one embodiment, each scFv domain has a structural order of N-terminus-VH-linker-VL-C-terminus or N-terminus-VL-linker-VH-C-terminus. In one embodiment, the linker comprises a flexible GS linker having about 20 amino acids. In one embodiment, the linker comprises the amino acid sequence (Gly-Gly-Gly-Gly-Ser)m, and wherein m is an integer of at least 3. In one embodiment, m is 4.

[0012] In one embodiment, the domain linker comprises from about 10 to about 30 amino acids. In one embodiment, the domain linker comprises the amino acid sequence (Gly-Gly-Gly-Gly-Ser)m, and wherein m is an integer of at least 2. In one embodiment, m is an integer from 2 to 6.

[0013] In one embodiment, the two epitopes of EGFR comprise an EGFR wild-type (EGFRwt) epitope and an EGFRvIII epitope, and the bispecific tetravalent antibody has a stronger binding affinity for EGFRvIII than for EGFRwt. In one embodiment, the antibody has a binding affinity of a first KD for the EGFRwt epitope and a binding affinity of a second KD for the EGFRvIII epitope, wherein the first KD is higher than the second KD.

[0014] In one embodiment, the first KD is not less than 1E-11 M and the second KD is not greater than 1E-06 M. In one embodiment, the first KD is between 1E-10 M and 1E-06 M, and the second KD is between 1E-11 M and 1E-07 M. In one embodiment, the first KD is 1.1-fold, 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold or 70-fold higher than the second KD. In one embodiment, the first KD is 1.1 to 1000-fold stronger than the second KD.

[0015] In one embodiment, the antibody has a first ADCC EC50 against cells carrying only EGFRwt and a second ADCC EC50 against cells carrying both EGFRwt and EGFRvIII, wherein the first ADCC EC50 is greater than the second ADCC EC50. In one embodiment, the first ADCC EC50 is 1.1-fold, 1.2-fold, 1.5-fold, 2-fold, 3-fold, 4-fold, or 5-fold higher than the second ADCC EC50.

[0016] In one embodiment, the EGFRwt epitope comprises an epitope having binding affinity for cetuximab. In one embodiment, the EGFRvIII epitope comprises an epitope having binding affinity for ABT-806.

[0017] In one embodiment, the scFv domain of the bispecific tetravalent antibody has binding affinity for the EGFRwt epitope and the Fab domain has binding affinity for the EGFRvIII epitope. In one embodiment, the scFv domain has binding affinity for the EGFRvIII epitope and the Fab domain has binding affinity for the EGFRwt epitope.

[0018] In one embodiment, the scFv domain is linked to the light chain of the bispecific tetravalent antibody at its C-terminus. In one embodiment, the scFv domain is linked to the light chain of the antibody at its N-terminus. In one embodiment, the scFv domain is linked to the heavy chain of the antibody at its C-terminus. In one embodiment, the scFv domain is linked to the heavy chain of the antibody at its N-terminus.

[0019] In one embodiment, the antibody backbone can be derived from cetuximab, humanized cetuximab, de-immunized cetuximab, or humanized de-immunized cetuximab (i.e., a cetuximab-derived backbone) or nimotuzumab, and the scFv domain can comprise a binding domain derived from ABT-806 or humanized ABT-806.

[0020] Alternatively, the antibody backbone can be derived from ABT-806 or humanized ABT-806, and the scFv domain can comprise a binding domain derived from cetuximab, humanized cetuximab, de-immunized cetuximab, or humanized de-immunized cetuximab or nimotuzumab.

[0021] In one embodiment, the humanized ABT-806 is ABT-806V1, V2, V3, V4, V5, V6, V7, V8, or V9.

[0022] Cetuximab backbone dual-epitope tetravalent antibody

[0023] In one embodiment, the present application provides a bispecific tetravalent antibody, wherein the antibody backbone is derived from cetuximab, humanized cetuximab, de - matured cetuximab or humanized de - matured cetuximab (i.e., a cetuximab - derived backbone). In one embodiment, the scFv domain comprises a binding domain derived from the variable regions of ABT - 806 or humanized ABT - 806.

[0024] In one embodiment, the antibody backbone comprises a cetuximab de - maturation mutation (numbered in sequence) selected from VH - Y101, VH - Y102, VH - D103, VH - Y104, VL - N92 or a combination thereof. In one embodiment, the cetuximab de - maturation mutation (numbered in sequence) comprises VH - Y101A, VH - Y101W, VH - Y102A, VH - D103F, VH - D103W, VH - D103Y, VH - Y104L, VL - N92F, VL - N92K or a combination thereof.

[0025] In one embodiment, the antibody heavy chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.1, 13, 79, 97, 101, 103, 105, 107, 123, 127, 129, 131 or 133. In one embodiment, the antibody light chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.3, 81, 83, 99, 109, 125 or 135.

[0026] In one embodiment, the antibody backbone comprises humanized cetuximab. In one embodiment, the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.237. In one embodiment, the antibody VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.239.

[0027] In one embodiment, the antibody backbone comprises de - matured cetuximab. In one embodiment, the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.247, 251, 253, 259, 261, 263, 265 or 267. In one embodiment, the antibody VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.243 or 269.

[0028] In one embodiment, the antibody backbone comprises humanized and de-matured cetuximab. In one embodiment, the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 245, 249, 255 or 257. In one embodiment, the antibody VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 241.

[0029] In one embodiment, the humanized ABT-806 is ABT-806V1, V2, V3, V4, V5, V6, V7, V8 or V9.

[0030] In one embodiment, the scFv domain comprises the variable regions of ABT-806 or humanized ABT-806.

[0031] In one embodiment, the scFv VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 209, 225, 229 or 233. In one embodiment, the scFv VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 211, 227, 231 or 235.

[0032] In one embodiment, the antibody VH domain comprises the 3 complementarity-determining regions (CDRs) of SEQ ID NO. 201, 237, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265 or 267. In one embodiment, the antibody VL domain comprises the 3 CDRs of SEQ ID NO. 203, 239, 241, 243 or 269. In one embodiment, the scFv VH domain comprises the 3 CDRs of SEQ ID NO. 209. In one embodiment, the scFv VL domain comprises the 3 CDRs of SEQ ID NO. 211.

[0033] In one embodiment, the bispecific tetravalent antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 1, 3, 13, 79, 81, 83, 97, 99, 101, 103, 105, 107, 109, 123, 125, 127, 129, 131, 133 or 135.

[0034] ABT-806 Framework Dual-Epitope Tetravalent Antibody

[0035] In one embodiment, the present application provides a dual-epitope tetravalent antibody, wherein the antibody framework is derived from ABT0806 or humanized ABT-806, and wherein the scFv domain comprises a binding domain derived from the variable region of cetuximab, humanized cetuximab, de-matured cetuximab, humanized de-matured cetuximab. In one embodiment, the scFv domain comprises a binding domain derived from nimotuzumab.

[0036] In one embodiment, the antibody heavy chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.9, 17, 19, 21, 23, 25, 27, 29, 31, 75, 93, 113, 115, 117, 119, 121, 137, 139, 141, 143, 145, 147 or 179.

[0037] In one embodiment, the antibody light chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.11, 33, 35, 37, 39, 41, 43, 77, 91, 95 or 111.

[0038] In one embodiment, the antibody framework comprises ABT-806, and wherein the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.209. In one embodiment, the VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.211.

[0039] In one embodiment, the antibody framework comprises humanized ABT-806. In one embodiment, the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.225, 229 or 233. In one embodiment, the antibody VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.227, 231 or 235.

[0040] In one embodiment, the scFv VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 201, 205, 237, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265 or 267.

[0041] In one embodiment, the scFv VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 203, 207, 239, 241, 243 or 269.

[0042] In one embodiment, the scFv domain comprises a binding domain derived from the variable region of a de-matured cetuximab or a humanized de-matured cetuximab. In one embodiment, the de-matured cetuximab or humanized de-matured cetuximab comprises a cetuximab de-maturation mutation selected from Y101, Y102, D103, Y104, N92 and combinations thereof (numbered in sequence). In one embodiment, the cetuximab de-maturation mutation (numbered in sequence) comprises VH-Y101, VH-Y102, VH-D103, VH-Y104, VL-N92 or combinations thereof. In one embodiment, the cetuximab de-maturation mutation (numbered in sequence) comprises VH-Y101A, VH-Y101W, VH-Y102A, VH-D103F, VH-D103W, VH-D103Y, VH-Y104L, VL-N92F, VL-N92K or combinations thereof.

[0043] In one embodiment, the VH domain of the bispecific tetravalent antibody comprises 3 complementarity determining regions (CDRs) of SEQ ID NO. 209. In one embodiment, the VL domain of the bispecific tetravalent antibody comprises 3 CDRs of SEQ ID NO. 211. In one embodiment, the scFv VH domain comprises 3 CDRs of SEQ ID NO. 201, 205, 237, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265 or 267. In one embodiment, the scFv VL domain comprises 3 CDRs of SEQ ID NO. 203, 207, 239, 241, 243 or 269.

[0044] In one embodiment, the bispecific tetravalent antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 9, 11, 17, 19, 21, 23, 25, 27, 29, 31, 33, 37, 41, 43, 75, 77, 91, 93, 95, 111, 113, 115, 117, 119, 121, 137, 139, 141, 143, 145, 147 or 179.

[0045] Nimotuzumab backbone antibody

[0046] In one embodiment, the present application provides a bispecific tetravalent antibody, wherein the antibody backbone comprises nimotuzumab, and wherein the scFv domain comprises a binding domain derived from the variable regions of ABT-806 or humanized ABT-806. In one embodiment, the scFv domain comprises the variable regions of ABT-806.

[0047] In one embodiment, the heavy chain of the antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 5, 15, 45, 47, 49, 51, 53, 55 or 85.

[0048] In one embodiment, the light chain of the antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 7, 57, 59, 61, 63, 65, 67, 69, 71 or 73.

[0049] In one embodiment, the VH domain of the antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 205.

[0050] In one embodiment, the VL domain of the antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 207.

[0051] In one embodiment, the scFv VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 209. In one embodiment, the scFv VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 211.

[0052] In one embodiment, the antibody VH domain comprises 3 complementarity-determining regions (CDRs) of SEQ ID NO. 205. In one embodiment, the antibody VL domain comprises 3 CDRs of SEQ ID NO. 207. In one embodiment, the scFv VH domain comprises 3 CDRs of SEQ ID NO. 209. In one embodiment, the scFv VL domain comprises 3 CDRs of SEQ ID NO. 211.

[0053] In one embodiment, the bispecific tetravalent antibody comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 5, 7, 15, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73 or 85.

[0054] Defucosylated cetuximab monoclonal antibody

[0055] In one aspect, the present application provides a de-matured cetuximab monoclonal antibody comprising a light chain having a light chain variable (VL) domain and a heavy chain having a heavy chain variable (VH) domain, wherein the VL domain and the VH domain form a Fab region, and wherein the monoclonal antibody comprises a cetuximab de-maturation mutation selected from VH-Y101, VH-Y102, VH-D103, VH-Y104, VL-N92 and combinations thereof (numbered in sequence).

[0056] In one embodiment, the cetuximab de-maturation mutation (numbered in sequence) comprises VH-Y101A, VH-Y101W, VH-Y102A, VH-D103F, VH-D103W, VH-D103Y, VH-Y104L, VL-N92F, VL-N92K or combinations thereof.

[0057] In one embodiment, the de-matured cetuximab monoclonal antibody is non-humanized.

[0058] In one embodiment, the antibody VH domain comprises 3 complementarity-determining regions (CDRs) having an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 247, 251, 253, 259, 261, 263, 265 or 267. In one embodiment, the antibody VL domain comprises 3 CDRs having an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 243 or 269.

[0059] In one embodiment, the heavy chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.1, 155, 157, 159, 161, 163, 165, 167 or 169. In one embodiment, the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.247, 251, 253, 259, 261, 263, 265 or 267.

[0060] In one embodiment, the light chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.3, 151 or 153. In one embodiment, the VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.243 or 269.

[0061] In one embodiment, the defucosylated cetuximab monoclonal antibody is humanized.

[0062] In one embodiment, the antibody VH domain comprises three complementarity determining regions (CDRs) having an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.245, 249, 255 or 257. In one embodiment, the antibody VL domain comprises three complementarity determining regions (CDRs) having an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.241.

[0063] In one embodiment, the heavy chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.123, 127, 129, 131 or 133. In one embodiment, the antibody VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.245, 249, 255 or 257.

[0064] In one embodiment, the light chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.99 or 109. In one embodiment, the antibody VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.241.

[0065] In one embodiment, the present application provides a multispecific antibody comprising an antibody backbone and at least one scFv domain linked to the antibody backbone, wherein the antibody backbone comprises a de-matured cetuximab monoclonal antibody as disclosed herein. In one embodiment, the present application provides a multispecific antibody comprising an antibody backbone and at least one scFv domain linked to the antibody backbone, wherein the scFv domain comprises the variable regions of a de-matured cetuximab monoclonal antibody as disclosed herein.

[0066] Humanized anti-EGFRvIII monoclonal antibody

[0067] In one aspect, the present application provides a humanized anti-EGFRvIII monoclonal antibody comprising a light chain having a light chain variable (VL) domain and a heavy chain having a heavy chain variable (VH) domain, wherein the VL domain and the VH domain form a Fab region. In one embodiment, the monoclonal antibody is ABT0806 or humanized ABT-806.

[0068] In one embodiment, the heavy chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.171, 149 or 177. In one embodiment, the light chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.111, 173 or 175. In one embodiment, the VH domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.225, 229 or 233. In one embodiment, the VL domain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO.227, 231 or 235.

[0069] In one embodiment, the present application provides a multispecific antibody comprising an antibody backbone and at least one scFv domain linked to the antibody backbone, wherein the antibody backbone comprises a humanized anti-EGFRvIII monoclonal antibody as disclosed herein. In one embodiment, the present application provides a multispecific antibody comprising an antibody backbone and at least one scFv domain linked to the antibody backbone, wherein the scFv comprises the variable regions of a humanized anti-EGFRvIII as disclosed herein.

[0070] Quadrivalent antibody

[0071] In one aspect, the present application provides a tetravalent antibody having binding affinity for EGFR, which comprises an antibody light chain having an antibody light chain variable (VL) domain, an antibody heavy chain having an antibody heavy chain variable (VH) domain, and an scFv domain having an scFv light chain variable (VL) domain and an scFv heavy chain variable (VH) domain, wherein the antibody VL domain and the antibody VH domain form a Fab region, and wherein the scFv domain is linked to at least one end of the antibody light chain or the antibody heavy chain.

[0072] In one embodiment, the antibody VH domain comprises an amino acid sequence having SEQ ID NO. 201. In one embodiment, the antibody VL domain comprises an amino acid sequence having SEQ ID NO. 203. In one embodiment, the antibody heavy chain comprises an amino acid sequence having SEQ ID NO. 89. In one embodiment, the antibody light chain comprises an amino acid sequence having SEQ ID NO. 3. In one embodiment, the scFv domain comprises a light chain variable (VL) domain having an amino acid sequence having SEQ ID NO. 203. In one embodiment, the scFv domain comprises a heavy chain variable (VH) domain having an amino acid sequence having SEQ ID NO. 201.

[0073] In one embodiment, the antibody heavy chain comprises an amino acid sequence having at least 80%, 85%, 90%, 95%, 98%, 99% sequence identity with SEQ ID NO. 89. In one embodiment, the antibody light chain comprises SEQ ID NO. 3. In one embodiment, the antibody VH comprises SEQ ID NO. 201. In one embodiment, the antibody VL comprises SEQ ID NO. 203. In one embodiment, the antibody scFv VH comprises SEQ ID NO. 201. In one embodiment, the antibody scFv VL comprises SEQ ID NO. 203.

[0074] In one embodiment, the antibody VH comprises 3 complementarity determining regions (CDRs) of SEQ ID NO. 201. In one embodiment, the antibody VL comprises 3 CDRs of SEQ ID NO. 203. In one embodiment, the antibody scFv VH comprises 3 CDRs: SEQ ID NO. 201. In one embodiment, the 3 CDRs of the antibody scFv VL comprise SEQ ID NO. 203.

[0075] In one embodiment, the tetravalent antibody is designated SI-95X42, as shown in the following examples.

[0076] In another embodiment, the present application provides a tetravalent antibody having binding affinity for EGFRvIII, which comprises an antibody light chain having an antibody light chain variable (VL) domain, an antibody heavy chain having an antibody heavy chain variable (VH) domain, and an scFv domain having an scFv light chain variable (VL) domain and an scFv heavy chain variable (VH) domain, wherein the antibody VL domain and the antibody VH domain form a Fab region, and wherein the scFv domain is linked to at least one end of the antibody light chain or the antibody heavy chain.

[0077] In one embodiment, the antibody VH domain comprises an amino acid sequence having SEQ ID NO. 209. In one embodiment, the antibody VL domain comprises an amino acid sequence having SEQ ID NO. 211.

[0078] In one embodiment, the antibody heavy chain comprises an amino acid sequence having SEQ ID NO. 87. In one embodiment, the antibody light chain comprises an amino acid sequence having SEQ ID NO. 11. In one embodiment, the scFv domain comprises a light chain variable (VL) domain having an amino acid sequence having SEQ ID NO. 211. In one embodiment, the scFv domain comprises a heavy chain variable (VH) domain having an amino acid sequence having SEQ ID NO. 209.

[0079] In one embodiment, the antibody heavy chain comprises SEQ ID NO. 87. In one embodiment, the antibody light chain comprises SEQ ID NO. 11. In one embodiment, the antibody VH domain comprises SEQ ID NO. 209. In one embodiment, the antibody VL domain comprises SEQ ID NO. 211. In one embodiment, the antibody scFv VH domain comprises SEQ ID NO. 209. In one embodiment, the antibody scFv VL domain comprises SEQ ID NO. 211.

[0080] In one embodiment, the antibody VH domain comprises 3 complementarity determining regions (CDRs) of SEQ ID NO. 209. In one embodiment, the antibody VL domain comprises 3 CDRs of SEQ ID NO. 211. In one embodiment, the antibody scFv VH domain comprises 3 CDRs of SEQ ID NO. 209. In one embodiment, the antibody scFv VL domain comprises 3 CDRs of SEQ ID NO. 211.

[0081] In one embodiment, the in-species antibody is represented by SI-95X41, as shown in the following examples.

[0082] In another aspect, the present application provides an isolated nucleic acid sequence encoding a bispecific tetravalent antibody, monoclonal antibody, tetra-specific antibody or multi-specific antibody as disclosed herein.

[0083] In yet another aspect, the present application provides an expression vector comprising a nucleic acid sequence as disclosed herein, wherein the vector is expressed in a cell.

[0084] In yet another aspect, the present application provides a host cell comprising a nucleic acid as disclosed herein. In one embodiment, the host cell comprises an expression vector as disclosed herein. In one embodiment, the host cell is a prokaryotic cell or a eukaryotic cell.

[0085] In yet another aspect, the present application provides a method for producing an antibody as disclosed herein. In one embodiment, the method comprises culturing a host cell as disclosed herein to produce the disclosed antibody. In one embodiment, the antibody is a bispecific tetravalent antibody. In one embodiment, the antibody is a monoclonal antibody. In one embodiment, the antibody is a tetra-specific antibody. In one embodiment, the antibody is a multi-specific antibody.

[0086] In yet another aspect, the present application provides an immunoconjugate comprising a bispecific tetravalent antibody as disclosed herein and a cytotoxic agent. In one embodiment, the cytotoxic agent comprises a chemotherapeutic agent, a growth inhibitor, a toxin or a radioisotope.

[0087] In yet another aspect, the present application provides a pharmaceutical composition comprising an antibody as disclosed herein and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition further comprises a radioisotope, a radionuclide, a toxin, a therapeutic agent, a chemotherapeutic agent or a combination thereof. In one embodiment, the antibody is a bispecific tetravalent antibody. In one embodiment, the antibody is a monoclonal antibody. In one embodiment, the antibody is a tetra-specific antibody. In one embodiment, the antibody is a multi-specific antibody.

[0088] In yet another aspect, the present application provides a pharmaceutical composition comprising an immunoconjugate as disclosed herein and a pharmaceutically acceptable carrier.

[0089] In yet another aspect, the present application provides a method for treating a subject suffering from cancer, which comprises administering to the subject an effective amount of a bispecific tetravalent antibody as disclosed herein. In one embodiment, the cancer comprises cells expressing EGFR, EGFRvIII or both. In one embodiment, the cancer includes breast cancer, colorectal cancer, pancreatic cancer, head and neck cancer, melanoma, ovarian cancer, prostate cancer, non-small cell lung cancer, small cell lung cancer, glioma, esophageal cancer, nasopharyngeal cancer, renal cancer, gastric cancer, liver cancer, bladder cancer, cervical cancer, brain cancer, lymphoma, leukemia, myeloma.

[0090] In one embodiment, the treatment method further comprises co-administering an effective amount of a therapeutic agent. In one embodiment, the therapeutic agent comprises an antibody, a chemotherapeutic agent, an enzyme, or a combination thereof. In one embodiment, the therapeutic agent comprises capecitabine, cisplatin, trastuzumab, fulvestrant, tamoxifen, letrozole, exemestane, anastrozole, aminoglutethimide, testolactone, vorozole, formestane, fadrozole, letrozole, erlotinib, lapatinib, dasatinib, gefitinib, imatinib, pazopanib, lapatinib, sunitinib, nilotinib, sorafenib, nab-paclitaxel, its derivatives, or combinations.

[0091] In one embodiment, the subject is a human.

[0092] In yet another aspect, the present application provides a solution comprising an effective concentration of a bispecific tetravalent antibody as disclosed herein, wherein the solution is the plasma of a subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0093] In conjunction with the drawings, the foregoing and other features of the present disclosure may become more apparent from the following description and the appended claims. It should be understood that these drawings only depict several embodiments arranged in accordance with the present disclosure and should not be considered as limiting its scope. The present disclosure can be described with additional specificity and detail by using the drawings, wherein:

[0094] Figure 1 Depicts the configuration of a bispecific tetravalent antibody capable of targeting two epitopes of EGFR, the bispecific tetravalent antibody comprising two anti-EGFR Fab domains and two anti-EGFRvIII scFv domains (A) or two anti-EGFRvIIIFab domains and two anti-EGFR scFv domains (B), wherein the scFv domains can be linked to the N-terminus or C-terminus of the heavy or light chain of the antibody, and the binding domains can be derived from cetuximab (which can be humanized and / or de-matured) or a combination of nimotuzumab and ABT-806 (which can be humanized);

[0095] Figure 2 Depicts a structural alignment of the complexes of cetuximab (PDB 1YY9), panitumumab (PDB 5SX4), and necitumumab (PDB6B3S) with EGFR or EGFR domain III, indicating that all three antibodies bind to overlapping epitopes (EGFRwt) within EGFR domain III;

[0096] Figure 3 shows the results of mass analysis of the wild-type and humanized forms of ABT806, where the wild-type contains two purified fractions corresponding to the intact mAb and the heavy-chain dimer contaminant (3A); and the two humanized forms show a single pure fraction containing the intact mAb of interest but lacking the heavy-chain dimer contaminant (3B);

[0097] Figure 4 shows the results of an ADCC assay measuring the potency of SI-95m4 (4A) and SI-95m5 (4B) against U87MG-EGFRwt(GFP) cells expressing EGFRwt and U87MG-EGFRvIII(NR) cells expressing EGFRwt / EGFRvIII (as separate or mixed target cells (T)) in the presence of NK effector cells (E:T = 5:1);

[0098] Figure 5 shows the results of an ADCC assay measuring the potency of a panel of anti-EGFRwt monoclonal antibodies (mAbs), including the de-matured variants SI-95m6, SI-95m7, SI-95m8, SI-95m9, SI-95m10, and controls of anti-EGFRvIII and anti-EGFRwt antibodies against U87MG-EGFRwt(GFP) cells expressing EGFRwt (5A) or U87MG-EGFRvIII(NR) cells expressing EGFRwt / EGFRvIII (5B) in the presence of NK effector cells (E:T = 5:1), and lists the EC50 values of the antibodies;

[0099] Figure 6 shows the results of an ADCC assay measuring the potency of a panel of bispecific tetravalent antibodies targeting EGFRwt and EGFRvIII (i.e., bispecific antibodies), including SI-95X44 and its de-matured derivatives, including but not limited to SI-95X52, SI-95X53, SI-95X54, SI-95X55, and SI-95X56, against U87MG-EGFRwt(GFP) cells expressing EGFRwt (6A) or U87MG-EGFRvIII(NR) cells expressing EGFRwt / EGFRvIII (6B) in the presence of NK effector cells (E:T = 5:1), and lists the EC50 values of the antibodies;

[0100] Figure 7 depicts the effect of the de-matured cetuximab mutants VH-Y101A (7A) or VH-Y102A (7B) on the bispecific tetravalent antibody (also known as bispecific antibody) SI-95X44 targeting both EGFRwt and EGFRvIII, compared to SI-95X65 and SI-95X66 respectively, in the presence of NK effector cells (E:T = 5:1), and the effect of having the humanized anti-EGFRvIII antibody ABT-806 backbone in SI-95X53 and SI-95X54, as measured by their potency against U87MG-EGFRwt(GFP) cells expressing EGFRwt, which indicates that both SI-95X53 and SI-95X54 have more complete killing against EGFRwt-expressing cells and have better EC50 values as listed; and

[0101] Figure 8 depicts the potential increase in the therapeutic index obtained by measuring the ADCC activity of the bispecific tetravalent antibody (or simply bispecific antibody) targeting both EGFRwt and EGFRvIII against U87MG-EGFRwt-expressing cells and U87MG-EGFRwt / EGFRvIII-expressing cells as a single or mixed population, by introducing the de-matured mutation Y102A into the mature cetuximab binding domain, and shows that SI-95X44 with mature cetuximab binding mediates overlapping killing curves with similar ED50 values, while SI-95X54 with the Y102A mutation differentiates the killing of the mixed population from that of the individual populations mimicking glioblastoma and EGFRwt-expressing cells in patient normal tissues. Detailed Description

[0102] Since the expression of wild-type and mutant EGFR (i.e., overexpressing EGFRwt protein and EGFRvIII mutant protein respectively) is highly prevalent in several types of solid tumors, developing an effective therapeutic strategy that simultaneously targets both forms of EGFR remains an unsolved challenge. Such a strategy would involve a first specificity for EGFRvIII and a second specificity for the wild-type receptor, where the first specificity would allow for preferential targeting of the most malignant cells and the second specificity would allow for broad tumor targeting. Such bispecific antibodies would represent a new class of bispecific tetravalent antibodies characterized by dual binding specificity for two epitopes of the same EGFR target molecule.

[0103] To construct a bispecific tetravalent antibody capable of binding both wild-type and mutant EGFR proteins (preferably binding EGFRvIII), scFv domains were attached to different ends of the antibody heavy and light chains. As Figure 1 shown, the scFv targeting EGFRvIII was attached to the N-terminus or C-terminus of the antibody targeting EGFR (via the heavy chain or light chain)(Figure 1 A, 1B, 1C, and 1D). Alternatively, the scFv targeting EGFRwt is attached to the N-terminus or C-terminus (via the heavy or light chain) of the antibody targeting EGFRvIII. For stability and binding affinity, several parameters are optimized, including the length of the linker between domains and the orientation of the scFv VH / VL domains (Table 1).

[0104] This application relates to methods for preparing and using bispecific tetravalent antibodies, particularly bisepitope tetravalent antibodies.

[0105] The term "epitope" defines the antigenic region that binds to an antibody. The term "antibody" is used in its broadest sense and specifically encompasses single monoclonal antibodies (including agonist and antagonist antibodies), antibody compositions with multi-epitope specificity, and antibody fragments (e.g., Fab, F(ab’)2, and Fv), provided they exhibit the desired biological activity. In some embodiments, the antibody can be a monoclonal antibody, polyclonal antibody, chimeric antibody, single-chain antibody, bispecific antibody or bifunctional antibody, human antibody, and humanized antibody, as well as active fragments thereof. Examples of active fragments of molecules that bind to known antigens include Fab, F(ab’)2, scFv, and Fv fragments, including products of Fab immunoglobulin expression libraries and epitope-binding fragments of any of the above antibodies and fragments. In some embodiments, the antibody can include an immunoglobulin molecule and the immunologically active portion of the immunoglobulin molecule, i.e., the molecule that contains the binding site and immunologically specifically binds to an antigen. The immunoglobulin can be any type (IgG, IgM, IgD, IgE, IgA, and IgY) or class (IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) or subclass of immunoglobulin molecule. In one embodiment, the antibody can be a whole antibody and any antigen-binding fragment derived from the whole antibody. A typical antibody refers to a heterotetrameric protein that generally comprises two heavy (H) chains and two light (L) chains. Each heavy chain consists of a heavy chain variable domain (abbreviated as VH) and a heavy chain constant domain. Each light chain consists of a light chain variable domain (abbreviated as VL) and a light chain constant domain. The VH and VL regions can be further subdivided into domains of hypervariable complementarity-determining regions (CDRs) and more conserved regions called framework regions (FRs). Each variable domain (VH or VL) typically consists of three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The binding region that interacts with the antigen is present within the variable regions of the light and heavy chains.

[0106] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a substantially homogeneous population of antibodies, i.e., the individual antibodies comprising the population are identical except for possible minor variations that may occur naturally. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations that typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, the advantage of monoclonal antibodies is that they are synthesized by hybridoma cultures and are not contaminated with other immunoglobulins.

[0107] The modifier "monoclonal" denotes the character of an antibody as obtained from a substantially homogeneous population of antibodies and should not be construed as requiring that the antibody be made by any particular method. In the case of antibodies directed against more than one common antigenic site, the term "monoclonal antibody" refers to a group of monoclonal monospecific antibodies in an early stage of development. For example, monoclonal antibodies useful according to the present disclosure can be prepared by the hybridoma method first described by Kohler & Milstein, Nature, 256:495 (1975), or can be prepared by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). In the latter case, the modifier "monoclonal" of the term "monoclonal multispecific antibody" is often omitted, which is understandable.

[0108] Monoclonal antibodies can include "chimeric" antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical or homologous to the corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain is identical or homologous to the 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. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855

[1984] ).

[0109] Monoclonal antibodies can be generated using various methods, including murine hybridomas or phage display (see review in Siegel. Transfus. Clin. Biol. 9:15 - 22 (2002)), or by directly molecularly cloning antibodies from primary B cells (see Tiller. New Biotechnol. 28:453 - 7 (2011)). Antibody variable genes are isolated using recombinant DNA techniques and the resulting antibodies are recombinantly expressed and further screened for desired characteristics such as the ability to inhibit the binding of EGFR in its wild - type or mutant forms. This general method of antibody discovery is similar to the method described in Seeber et al. PLOS One. 9:e86184 (2014). Once monoclonal antibody genes are discovered, they can be humanized, engineered, combined, and otherwise modified to generate various types of multispecific antibodies. Cetuximab (anti - EGFR) and ABT - 806 (anti - EGFRvIII) were both discovered using murine hybridomas. 9,10

[0110] In the present application, the term "antigen or epitope - binding portion or fragment" refers to a fragment of an antibody that is capable of binding an antigen such as EGFR. These fragments may have the antigen - binding function of the full - length antibody and additional functions. Examples of binding fragments include, but are not limited to, single - chain Fv fragments (scFv), which consist of the VL and VH domains of a single arm of an antibody joined by a synthetic linker in a single polypeptide chain, or Fab fragments, which are monovalent fragments consisting of the VL, constant light chain (CL), VH, and constant heavy chain 1 (CH1) domains. Antibody fragments can even be smaller sub - fragments and can consist of domains as small as a single CDR domain, particularly the CDR3 region from the VL and / or VH domains (see, for example, Beiboer et al., J. Mol. Biol. 296:833 - 49 (2000)). Antibody fragments are generated using conventional methods known to those skilled in the art. The utility of antibody fragments can be screened using the same techniques employed for full - length antibodies.

[0111] "Antigen or epitope - binding fragments" can be derived from the antibodies of the present disclosure by several techniques known in the art. For example, purified monoclonal antibodies can be cleaved with an enzyme such as pepsin and subjected to HPLC gel filtration. Appropriate fractions containing Fab fragments can then be collected and concentrated by, for example, membrane filtration. For further description of general techniques for isolating antibody active fragments, see, for example, Khaw, B.A. et al. J. Nucl. Med. 23:1011 - 1019 (1982); Rousseaux et al. Methods Enzymology, 121:663 - 69, Academic Press, 1986.

[0112] Papain digestion of an antibody produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and also produces a residual "Fc" fragment, so named for its ability to crystallize readily. Pepsin treatment yields F(ab’)2 fragments that have two antigen-binding sites and are still capable of cross-linking antigens.

[0113] Fab fragments can contain the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab’ fragments differ from Fab fragments by the addition of several residues at the carboxyl terminus of the heavy chain CH1 domain, including one or more cysteines from the antibody hinge region. Fab’-SH refers herein to a Fab’ in which the cysteine residue of the constant domain bears a free thiol group. F(ab’)2 antibody fragments are initially produced as pairs of Fab’ fragments that have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0114] "Fv" is the smallest antibody fragment that contains a complete antigen-recognition and binding site. This region is a dimer of one heavy-chain variable domain and one light-chain variable domain that are tightly, non-covalently associated. In this configuration, the three CDRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. The six CDRs together confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three antigen-specific CDRs) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0115] Based on the amino acid sequence of the constant domain, the "light chain" of an antibody (immunoglobulin) from any vertebrate species can be assigned to one of two distinct types, called κ and λ.

[0116] Immunoglobulins can be classified into different classes based on the amino acid sequence of the constant domain of the heavy chain. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), e.g., IgG-1, IgG-2, IgG-3, and IgG-4; IgA-1 and IgA-2. The constant domains of the heavy chains corresponding to the different classes of immunoglobulins are designated α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known.

[0117] "Humanized antibody" refers to a class of engineered antibodies having CDRs derived from non-human donor immunoglobulins, with the remaining immunoglobulin-derived portions of the molecule derived from one (or more) human immunoglobulins. Additionally, framework support residues may be altered to maintain binding affinity. Methods for obtaining "humanized antibodies" are well known to those skilled in the art. (See, e.g., Queen et al., Proc. Natl Acad Sci USA, 86:10029-10032 (1989); Hodgson et al., Bio / Technology, 9:421 (1991)).

[0118] As used herein, the terms "polypeptide", "peptide" and "protein" are interchangeable and are defined to mean a biomolecule composed of amino acids linked by peptide bonds.

[0119] As used herein, the terms "a", "an" and "the" are defined to mean "one or more" and include the plural, unless the context is inappropriate.

[0120] "Isolated" means that a biomolecule is free of at least some of the components in which it naturally occurs. When used to describe the various polypeptides disclosed herein, "isolated" means a polypeptide that has been identified and separated and / or recovered from the cell or cell culture in which it was expressed. Typically, an isolated polypeptide will be prepared by at least one purification step. An "isolated antibody" is an antibody that is substantially free of other antibodies having different antigen-binding specificities.

[0121] "Recombinant" means the generation of an antibody in a foreign host cell using recombinant nucleic acid techniques.

[0122] The term "antigen" refers to an entity or a fragment thereof that can induce an immune response in an organism, particularly an animal, more particularly a mammal including a human. The term includes immunogens and the regions thereof responsible for antigenicity or antigenic determinants.

[0123] In addition, as used herein, the term "immunogenic" refers to a substance that initiates or enhances the production of antibodies, T cells or other reactive immune cells against an immunogenic agent and contributes to an immune response in a human or an animal. An immune response occurs when an individual generates sufficient antibodies, T cells and other reactive immune cells against the administered immunogenic composition of the present disclosure to mitigate or alleviate the disorder to be treated. In the case of preparing a humanized antibody, the term "immunogenic" refers to the ability of a therapeutic agent such as a non-humanized antibody to elicit an undesired immune response against the therapeutic agent.

[0124] "Specific binding", "specifically binds to", or "is specific for a particular antigen or epitope" means binding that is measurably different from non-specific interactions. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule, which is typically a molecule of similar structure that does not have binding activity. For example, specific binding can be determined by competition with a control molecule similar to the target.

[0125] Specific binding to a particular antigen or epitope can be manifested, for example, by an antibody having a KD for the antigen or epitope of at least about 10-4 M, at least about 10-5 M, at least about 10-6 M, at least about 10-7 M, at least about 10-8 M, at least about 10-9 M, alternatively at least about 10-10 M, at least about 10-11 M, at least about 10-12 M or greater, where KD refers to the dissociation rate of a particular antibody-antigen interaction. Typically, the KD of an antibody that specifically binds an antigen is 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5000-fold, 10000-fold or more that of a control molecule relative to the antigen or epitope.

[0126] In addition, specific binding to a particular antigen or epitope can be manifested, for example, by an antibody having a KA or Ka for the epitope that is at least 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5000-fold, 10000-fold or more that of the epitope relative to a control, where KA or Ka refers to the association rate of a particular antibody-antigen interaction.

[0127] "Homology" between two sequences is determined by sequence identity. If the two sequences to be compared are of different lengths, sequence identity preferably refers to the percentage of nucleotide residues of the shorter sequence that are identical to the nucleotide residues of the longer sequence. Sequence identity can be routinely determined using computer programs. Deviations that occur in the comparison between a given sequence and the above sequences of the present disclosure can be caused, for example, by addition, deletion, substitution, insertion or recombination.

[0128] The term "dematuration" refers to the process of engineering a mature antibody, such as cetuximab in the present application, to have mutations (such as point mutations) in its binding domain. The resulting antibody is called a "dematured" antibody, which exhibits altered but desired binding activity that is characteristically different from the binding activity of the "matured" antibody.

[0129] The term "ADCC" refers to antibody-dependent cell cytotoxicity, which is an adaptive immune response in which effector cells carrying Fc receptors recognize and kill antibody-coated target cells that express tumor-associated or pathogen-derived antigens on their surface. ADCC is widely used to characterize and compare therapeutic candidates of monoclonal antibodies. Classical ADCC is mediated by natural killer (NK) cells.

[0130] The present disclosure can be more easily understood by referring to the following detailed description of specific embodiments and examples included herein. Although the present disclosure has been described with reference to the specific details of certain embodiments, such details are not intended to limit the scope of the present disclosure.

[0131] Example

[0132] Example 1. Engineered dual-epitope tetravalent antibody targeting EGFR

[0133] Using standard molecular biology techniques, genes encoding the heavy and light chains of the antibody (preceded by Kozak and a secretion signal peptide) were cloned into the pTT5 vector. Antibodies and bispecific antibodies were expressed by transient transfection of the expression plasmids for the heavy and light chains in the ExpiCHO system (Thermo Fisher). Briefly, 10 μg of each expression plasmid was added to 1 ml with OptiPRO SFM medium. 1 ml of OptiPRO SFM medium containing 80 μl of Expifectamine CHO reagent was added to the DNA and incubated at room temperature for 2.5 minutes. Then the resulting mixture was added to 25 ml of ExpiCHO cells in a 125-ml Erlenmeyer flask at 6×106 cells / ml and incubated at 37 °C, 5% CO2, 150 rpm. 24 hours after transfection, the cells were given 8.75 ml of ExpiCHO Feed and 150 μl of CHO Enhancer, and the temperature was changed to 32 °C, 5% CO2, 150 rpm. 48 hours after transfection, the cells were given 8.75 ml of ExpiCHO Feed again. 9 days after transfection, the culture supernatant was harvested, centrifuged at 4500 rpm for 1 hour to pellet the cells, and then passed through a 0.2-μm filter. The expression titer was quantified using a protein A sensor on an Octet384 system and a standard curve prepared from purified bispecific antibody protein by biolayer interferometry.

[0134] On an AKTA Avant system, the protein was purified from the harvested supernatant using a 5-ml MabSelect PrismA protein A column (GE Healthcare) equilibrated in phosphate-buffered saline. Then the supernatant was passed through the column at a flow rate of 5 ml / min and washed with 25 ml of PBS (125 mM sodium phosphate, 137 mM sodium chloride, pH 6.8). Then 15 ml of 50 mM sodium acetate (pH 3.5) was passed through the column to elute the protein of interest. The eluted protein was immediately neutralized by adding 0.5 ml of 1 M Tris-Cl (pH 8.0).

[0135] After the first step of Protein A purification, immediately use ACQUITY controlled by Waters Acquity UPLC H-Class Protein BEH SEC A 4.6 mm × 150 mm, 1.7 μm column was used to analyze the protein by analytical SEC. 10 μg of the protein was injected into the mobile phase of PBS and run for 10 minutes at 0.3 ml / min. On an AKTA Pure system, the protein was further purified by preparative SEC using a Superdex Increase 10 / 300Gl column in a mobile phase of 25 mM sodium acetate, 125 mM NaCl, pH 5.5, and finally buffer-exchanged into 25 mM sodium acetate, 125 mM NaCl, 10% sucrose, pH 5.5. The final sample contained >95% of the protein of interest (POI) as evaluated by analytical SEC and was used for subsequent assays.

[0136] Biolayer interferometry (Octet) binding assays (affinity) were performed on an Octet384 instrument to quantify the binding kinetics of the bispecific antibody to EGFR and EGFRvIII. The antibody was captured onto an anti-human Fc (AHC) sensor tip by loading at 5 μg / ml for 180 s. After a 60 s baseline step, an 180 s association period with serially diluted (0 - 100 nM; 1:2 dilution factor) His-tagged EGFR or EGFRvIII (internally expressed and purified) in assay buffer (PBS containing 0.1% BSA, 0.05% Tween 20) was carried out, followed by a 300 s dissociation period in assay buffer solution. Regeneration was achieved using 10 mM glycine (pH 1.5). The binding curves were globally fit to a 1:1 model to obtain the dissociation constant KD and the kinetic association and dissociation rates.

[0137] Biolayer interferometry (Octet) binding assays (avidity) were performed on an Octet384 instrument to quantify the divalent binding kinetics of the bispecific antibody to immobilized EGFR and EGFRvIII. Biotinylated EGFR or EGFRvIII was captured onto streptavidin (SA) sensor tips by loading at 1 μg / ml for 120 s. After a 60 s baseline step, an 180 s association period with serially diluted (0 - 25 nM; 1:2 dilution factor) antibody in assay buffer was carried out, followed by a 300 s dissociation period in assay buffer solution. Regeneration was achieved using 10 mM glycine (pH 1.5). The binding curves were globally fit to a 1:1 model to obtain the dissociation constant KD and the kinetic association and dissociation rates.

[0138] Epitope grouping studies were performed using biolayer interferometry (Octet). Biotinylated EGFR-WT (AcroEGR-H82E3) or EGFRvIII (Acro EGR-H82E0) was immobilized on streptavidin sensors in assay buffer (PBS containing 0.1% BSA, 0.05% Tween 20) at 1 μg / ml for 180 seconds. After a 30-second baseline, association with 100 nM of the first antibody was carried out for 300 seconds. Finally, association with 100 nM of the second antibody was carried out for 300 seconds. An epitope grouping matrix was generated using Octet analysis 12.0 software.

[0139] Example 2: Expression titer

[0140] Protein stability is a key parameter defined by the free energy difference between the folded and unfolded states. For protein therapeutics, stability can affect immunogenicity, pharmacokinetics, and even efficacy. Reducing aggregation can help develop therapeutics that are easier to manufacture and safer for patients. Additionally, expression efficiency and protein yield directly determine the cost of protein therapeutics. If proteins can be expressed effectively to achieve higher titers and increased yields of purified proteins, the manufacturing cost can be significantly reduced.

[0141] After transient expression in ExpiCHO cells, titers were quantified using biolayer interferometry (Table 2). Monoclonal antibodies with simple structures (EGFR or EGFRvIII) were produced with the highest efficiency, while bispecific (EGFR×EGFRvIII, SI-95X1-31, SI-95X35-39) antibodies were produced with variable efficiency, mainly depending on the composition of the binding domains. Bispecific antibodies with nimotuzumab scFv (SI-95X4-16) were produced at significantly lower titers, indicating instability of the VH and VL of nimotuzumab in this scFv form. On the other hand, constructs with nimotuzumab at the Fab position of the antibody and ABT-806 scFv (SI-95X2, SI-95X17-31) tended to show on average higher titers.

[0142] Example 3: Stability determined by analytical size exclusion chromatography

[0143] After the first step of Protein A purification, the stability and aggregation of the bispecific EGFR×EGFRvIII antibody were immediately evaluated by analytical size-exclusion chromatography on a Waters UPLC system (Table 2). Antibodies containing the nimotuzumab scFv (SI-95X4-16) typically showed low signals due to low expression and often had high abundances of high molecular weight (HMW) or low molecular weight (LMW) contaminants in addition to the main peak corresponding to the protein of interest (POI). This indicates that the nimotuzumab scFv is unstable when incorporated into the bispecific antibody. The other bispecific antibodies on average showed more protein of interest and less HMW and LMW species. The protein with the C-terminal scFv fused to the heavy chain (SI-95X1-3) showed the highest %POI, indicating good stability. The scFv orientation also seems to affect the SEC profile, with antibodies containing the ABT-806 scFv showing a complex split profile when in the HL orientation (SI-95X17-19, SI-95X23-25, SI-95X29-31), but no corresponding structure when the ABT-806 scFv is in the LH orientation (SI-95X20-22, SI-95X26-28). This result suggests that the LH orientation of the scFv may be more stable and / or conformationally homogeneous. The effect of the domain linker length on aggregation was negligible.

[0144] Example 4: Octet binding to EGFR WT (EGFRwt)

[0145] Biolayer interferometry was used to evaluate the affinity of the bispecific antibodies for wild-type EGFR (Table 3). The cetuximab and nimotuzumab antibodies showed strong binding to wild-type EGFR, as demonstrated by response values of 0.75 and 0.32 nm, respectively, while ABT-806, an EGFRvIII-specific antibody, showed a much lower response of 0.07 nm. Most of the EGFR×EGFRvIII bispecific antibodies showed strong binding to wild-type EGFR, except for those containing the nimotuzumab scFv and ABT-806 Fab (SI-95X4-16) which showed low binding. This result indicates that the function of nimotuzumab is reduced when present in the scFv form. On the other hand, good binding to wild-type EGFR was observed when nimotuzumab was in the Fab position (SI-95X2, SI-95X17-31), or when cetuximab was in the Fab (SI-95X1, SI-95X37-39) or scFv (SI-95X3, SI-95X35-36) position. The affinity data showed that antibodies with the cetuximab domain had a stronger affinity (lower KD) than those with the nimotuzumab domain.

[0146] Example 5: Octet binding to EGFRvIII

[0147] Biolayer interferometry was used to evaluate the affinity of bispecific antibodies for EGFRvIII (Table 4). As expected, all monoclonal and bispecific antibodies targeting wild-type EGFR and / or EGFRvIII showed detectable binding to EGFRvIII, which is consistent with the presence of two binding epitopes in the EGFRvIII protein. Cetuximab had a stronger binding response and affinity compared to nimotuzumab, while the binding of ABT-806 was intermediate between the two. For the bispecific antibody (SI-95X4-16) containing the nimotuzumab scFv and the ABT-806 Fab, the binding observed was similar to that of the ABT-806 monoclonal antibody, indicating that the nimotuzumab scFv did not contribute additional functionality. In contrast, the bispecific antibodies (SI-95X2, SI-95X17-31) containing the nimotuzumab Fab and the ABT-806 scFv had lower KD values and slower dissociation rates and, in some cases, showed enhanced affinity compared to the component mAbs. The same affinity enhancement was observed for the bispecific antibodies (SI-95X1, SI-95X3, SI-95X35-39) containing cetuximab and the ABT-806 domain. The results suggest that the nimotuzumab / cetuximab and ABT-806 domains can simultaneously bind two epitopes of the same EGFRvIII molecule, resulting in a multivalent avidity effect that enhances the apparent affinity.

[0148] Example 6: Epitope grouping (EGFRwt epitope)

[0149] Biolayer interferometry was used to epitope-map a panel of monoclonal antibodies using wild-type EGFR (Table 5). The antibodies included the EGFR-specific cetuximab (Cetu), panitumumab (Pani), nimotuzumab (Nimo), and necitumumab (Neci), as well as the EGFRvIII-specific ABT-806. Larger response values indicate that the second antibody was able to associate with EGFR even after saturation with the first antibody (indicating different epitopes). In contrast, smaller values indicate that the second antibody was blocked by the first antibody through competitive inhibition (indicating the same epitope).

[0150] As expected, ABT-806 always showed a low response as the second antibody because ABT-806 does not appreciably bind wild-type EGFR. All EGFR-targeting antibodies showed a high response as the second antibody after ABT-806 association because ABT-806 was unable to bind and block the second antibody.

[0151] In contrast, each of the four EGFR-targeting antibodies blocks the others to some extent, indicating identical or overlapping epitopes. Notably, if nimotuzumab is used as the first antibody, the four EGFR-targeting antibodies have detectable but reduced binding. This may be because nimotuzumab has slow association kinetics and does not fully saturate EGFR during the initial 300-second step. In summary, the data confirm that ABT-806 does not significantly bind to wild-type EGFR and suggest that all four EGFR-targeting antibodies (cetuximab, panitumumab, nimotuzumab, necitumumab) share a conserved epitope. This latter conclusion is supported by other studies that have shown that panitumumab, cetuximab, and nimotuzumab have overlapping epitopes within EGFR domain III. The crystal structures of EGFR complexed with the Fabs of cetuximab (1YY9), panitumumab (5SX4), and necitumumab (6B3S) were structurally aligned to compare the epitopes of these EGFR-targeting antibodies using the MatchMaker function in Chimera 1.13( Figure 2 ). This alignment demonstrated that all three antibodies bind to overlapping epitopes within EGFR domain III, consistent with the epitope grouping data from Octet. 11-12

[0152] Example 7: Epitope grouping (EGFRvIII epitope)

[0153] Using biolayer interferometry, the same set of monoclonal antibodies was grouped by epitope using EGFRvIII (Table 6). Larger response values indicate that the second antibody is still able to associate with EGFR even after saturation with the first antibody (indicating different epitopes). In contrast, smaller values indicate that the second antibody is blocked by the first antibody through competitive inhibition (indicating identical epitopes).

[0154] As observed in the wild-type EGFR grouping experiment, all four EGFR-targeting antibodies (cetuximab, panitumumab, nimotuzumab, and necitumumab) showed competitive binding to each other, as demonstrated by the reduced binding of the second antibody after saturation with the first antibody. This indicates that these four antibodies share overlapping epitopes on EGFRvIII, which may not be EGFRvIII-specific, i.e., the EGFRwt epitope. When nimotuzumab was used as the first (blocking) antibody, a slightly higher response of the second antibody was observed, which can be explained by the fact that nimotuzumab did not saturate the EGFRvIII protein during the initial 300-second blocking step due to its slower association rate.

[0155] In contrast, unlike the four EGFR-targeting antibodies, ABT-806 clearly groups to a different epitope, namely, the EGFRvIII-specific epitope. When ABT-806 is used as the first antibody, each of the other antibodies shows a significant association response in the second step. Similarly, when ABT-806 is used as the second antibody, a large additional response is observed. The lack of competitive binding between ABT-806 and the other antibodies indicates that ABT-806 targets a unique epitope on EGFRvIII, and that ABT-806 and other antibodies that bind to EGFR can simultaneously interact with different epitopes on the EGFRvIII protein. Compared to the EGFR-targeting antibodies, ABT-806 binds to a different epitope of EGFRvIII, which is consistent with the conformation of ABT-806 binding to the EGFR protein, which is limited to the mutant EGFRvIII and absent on the wild-type EGFR.

[0156] Example 8: Generation and characterization of humanized ABT-806

[0157] To increase the human nature of the ABT-806 variable regions and reduce the potential for immunogenicity, the murine VH and VK domains were converted to more human frameworks. All forms used an scFv model generated by the antibody modeling feature of DiscoveryStudio based on the sequence of the ABT-806 variable domain. All humanized forms were designed using the Discovery Studio 2022 suite. VH1, VH2, VK1, and VK2 were designed using the "Predict Humanizing Mutations" protocol, where the identity threshold was set to 50, the common residue substitution tolerance was set to 20, the germline substitution tolerance was set to 0, and substitutions of Kabat CDR residues, IMGT CDR residues, Vernier region residues, and human germline residues were excluded. This protocol was based solely on the amino acid sequence of ABT-806 as the query sequence. VH1 and VK1 were generated based on germline substitutions, while VH2 and VK2 forms used common residue substitutions. VH3 and VK3 were designed using the input Fv model of ABT806, where "Calculate Mutation Energy" was set to true (CHARMm force field) in order to generate the "best single mutation" sequence. The query structure was a model of ABT-806 generated by the Antibody Modeling Cascade of Discovery Studio. The input sequences were ABT-806 VH and VL. The CDR loop definition was set to Honegger, the maximum template for each loop was set to 3, and the optimization level was set to high.

[0158] Each humanized chain was combined to generate 9 forms: V1 (VH1, VK1), V2 (VH1, VK2), V3 (VH1, VK3), V4 (VH2, VK1), V5 (VH2, VK2), V6 (VH2, VK3), V7 (VH3, VK1), V8 (VH3, VK2), V9 (VH3, VK3) (Table 7). The heavy and light chain genes were cloned and expressed as in Example 1, and then the stability and functional properties of the antibodies were evaluated.

[0159] After transient expression in ExpiCHO cells, the titers were quantified using biolayer interferometry (Table 7). The best humanized forms were selected (partially) based on the overall expression.

[0160] As described in Example 1, the antibodies were purified by Protein A and SEC chromatography. After Protein A purification, in addition to the main peak corresponding to the protein of interest (POI), many antibodies often also had high-abundance high molecular weight (HMW) or low molecular weight (LMW) contaminants. This indicates that these antibodies are less stable, more prone to forming other substances (HC dimers), and / or more likely to aggregate. Based on the above metrics, V3 and V9 seem to be the most stable humanized ones (Table 7). The samples for subsequent studies were purified to >95% purity by preparative SEC.

[0161] As described in Example 1, biolayer interferometry (Octet) binding assays were performed on an Octet384 instrument to quantify the binding kinetics of the humanized antibodies to EGFR and EGFRvIII. Table 8 shows that all monoclonal antibodies targeting EGFRvIII exhibited detectable binding to EGFRvIII. Although all antibodies showed similar binding kinetics, the humanized V3 and V9 antibodies seemed to have a higher response, comparable to ABT-806 and superior to the competitor ditumumab, i.e., the humanized ABT-806 (internally purified) from AbbVie.

[0162] To confirm the binding of the antibodies to cells, the purified primary antibodies were added to CHO cells transfected with EGFR or EGFRvIII or uPAR. A fluorescent secondary antibody against the primary antibody was used to measure the antigen binding at the cell surface. All tested antibodies retained their binding properties to the cells mimicking the cancer surface (Table 9). Additionally, the binding of all antibodies to CHO cells expressing EGFRvIII was significantly better than that to CHO cells expressing EGFR. This demonstrated that these antibodies were more prone to binding to the ABT-806 epitope rather than the cetuximab epitope.

[0163] Preparative SEC (pSEC) was used prior to the complete mass analysis in order to partially separate the full mAb molecules (pSEC fraction 1) from the heavy chain dimer molecules (pSEC fraction 2). For quality analysis and confirmation of the pSEC fractions, the samples were diluted to 0.5 mg / mL with 50 mM tris pH 7.5. 1.0 μg of the prepared sample was injected onto an online analytical SEC column for LC-MS complete mass analysis under denaturing conditions. The data was processed using the MaxEnt 1 algorithm in Unifi (Waters Corp). As Figure 3A shown, pSEC fraction 1 of ditumumab contained almost only intact full mAb molecules, while the subsequently eluted pSEC fraction 2 contained a mixture of full mAb and heavy chain dimers. Figure 3B It was shown that both the purified hABT-806 V3 and V9 antibodies contained almost all intact full mAbs. Thus, the humanization of ABT806 generated more stable antibodies and had fewer problematic contaminants expressed.

[0164] The variable domains of the leader heavy and light chains were compared for sequence similarity with the LENS database. The results showed that the variable region sequences of VH3, VK3, and VH1 were unique (Table 10).

[0165] Example 9. Generation and Characterization of De-matured Cetuximab

[0166] A computer simulation (in silico)-based method was used to identify the mutations in cetuximab that de-matured its binding to EGFR. Using the Analyze Protein Interface Method in Discovery Studio (Biovia), the contact area between cetuximab and EGFR was calculated based on the high-resolution crystal structure of cetuximab binding to EGFR (1YY9). Nine contact surface areas were greater than Cetuximab residues were identified as strongly involved in binding to EGFR. The residues of interest are as follows (numbered in order): light chain W94, N91, and N92; heavy chain W52, N56, Y101, Y102, D103, Y104. Each residue of interest was mutated to one of 20 naturally occurring amino acids separately. A total of 180 (9 residues × 20 amino acids) independent mutations were generated by computer simulation. The Discovery Studio Calculate MutationEnergy (Stability) function was used to calculate the effect of the mutations on the stability of cetuximab, while the CalculateMutation Energy (Binding) function was used to calculate the effect of the mutations on the binding to EGFR, where EGFR was set as the ligand molecule. Briefly, the stability function calculates the energy difference (ΔΔG stability) between WT cetuximab and the mutant cetuximab, while the binding function calculates the binding energy between cetuximab and EGFR (ΔΔG WT), and then calculates the binding energy between the cetuximab mutant and EGFR (ΔΔG mutant), where the difference between the energies is reported as the energy change of binding due to the mutation (ΔΔ binding = ΔΔG mutant - ΔΔG WT). CHARMm was used to calculate the energies in a pH-independent mode. For all mutations, ΔΔG stability was plotted against the corresponding ΔΔG binding to identify a series of mutations that might cause binding de-maturation and that would not strongly destabilize cetuximab. Ten mutations were selected for further analysis: light chain N92K, N92F; heavy chain Y101A, Y101W, Y102V, Y102A, D103W, D103Y, D103F, and Y104L (Table 11).

[0167] The following describes the preparation of the de-matured cetuximab variants. Using standard molecular biology techniques, the codon-optimized coding region preceded by Kozak and a secretion signal peptide sequence was cloned into the pTT5 vector. As described in Example 1, the antibody was expressed by transient transfection of the light and heavy chains in the ExpiCHO system (ThermoFischer Scientific), but scaled down to a 2 mL culture volume in a 6-well plate. The protein was purified from the harvested supernatant using Captureem Protein A 24-well plates (TaKaRa). The supernatant was loaded onto plates equilibrated with phosphate-buffered saline (PBS, 125 mM sodium phosphate, 137 mM sodium chloride, pH 6.8). The plates were washed with 10 mL of PBS and then the bound protein was eluted with 0.5 mL of 50 mM sodium acetate (pH 3.5). The eluted protein was immediately neutralized by adding 0.1 mL of 1 M Tris-Cl, pH 8.0. Immediately after Protein A purification, the protein was analyzed by analytical SEC as described in Example 1. The Biolayer Interferometry (Octet) binding assay was performed as described in Example 1.

[0168] To evaluate the stability of the de-matured cetuximab variants, Biolayer Interferometry and analytical size exclusion chromatography were used. After transient expression in ExpiCHO cells, the titers were quantified using Biolayer Interferometry (Table 12). The selection of the optimal de-matured form was to some extent determined by the overall expression. Immediately after the first Protein A purification, the stability and aggregation of the antibody were evaluated by analytical size exclusion chromatography on a Waters UPLC system. In addition to the main peak corresponding to the protein of interest (POI), many antibodies often also have high molecular weight (HMW) species (Table 12). A higher abundance of HMW species indicates that these antibodies are less stable and more prone to aggregation.

[0169] The affinity and avidity of the de-matured cetuximab for EGFR and EGFRvIII were evaluated using Biolayer Interferometry, respectively. Table 13 shows that compared to the wild-type cetuximab, most de-matured cetuximab variants have a reduced binding affinity and / or reduced binding response to EGFR, indicating that the selected mutations altered the ability of cetuximab to bind EGFR. However, the antibody retained good avidity for immobilized EGFR.

[0170] Table 14 shows that most of the de-matured cetuximab variants also have reduced binding affinity for EGFRvIII and / or reduced binding response. Despite the reduction, all of the selected de-matured cetuximab variants still retain the ability to bind to EGFRvIII, with similar binding kinetics. This data again indicates that cetuximab binds to the same epitope on EGFRvIII and EGFR. Despite the weaker affinity of the variants for EGFR and EGFRvIII, due to bivalent binding, they maintain good avidity for EGFR and EGFRvIII. Since avidity depends on the antigen level on the surface, this result suggests that the de-matured form of cetuximab may be more selective for tumors carrying high amounts of EGFR and / or EGFRvIII.

[0171] To confirm that the de-matured cetuximab variants retained the ability to bind to cells, purified primary antibodies were added to CHO cells transfected with EGFR, EGFRvIII, or uPAR. Fluorescent secondary antibodies against the primary antibodies were used to measure antigen binding at the cell surface. Table 15 shows that all of the tested antibodies (except for HC:D103W) retained their binding properties to cells mimicking the cancer surface. The results indicate that these antibodies retained the binding behavior required for preparing antibody therapeutics.

[0172] Example 10. Generation and characterization of dual-epitope tetravalent antibody targeting EGFR

[0173] After the approval of cetuximab, nimotuzumab has been approved in several countries for the treatment of solid tumors. At least one of its improvements is reduced skin toxicity, which seems to be related to its lower affinity for EGFR, i.e., a KD value of 9.1 nM compared to 3.77 nM (Table 3).9 In this case, the most kinetically comparable cetuximab variant is Y104L (KD 9.2 nM), while all other variants exhibit even weaker EGFR affinity than nimotuzumab, including Y101A (KD 67 nM), Y102A (97 nM), and N92F (34 nM). These de-matured variants were incorporated into bispecific tetravalent antibodies to screen for potential therapeutic candidates that effectively target tumors expressing EGFRvIII in the presence of EGFRwt.

[0174] Bispecific tetravalent antibodies targeting EGFR and EGFRvIII, also known as anti-EGFR×EGFRvIII antibodies, were constructed by fusing scFv to the N-terminus or C-terminus of the heavy or light chain of the antibody. Figure 1 Four exemplary structures are depicted, where the EGFR-binding domain can be derived from humanized and / or de-matured cetuximab variants (hDC-EGFR) and are formatted as Fab or scFv, while ABT-806 or humanized ABT-806 can be conversely formatted as scFv or Fab.

[0175] Table 16 lists four sets of configurations of anti-hDC-EGFR×hABT-806 antibodies, namely SI-95X45-49, SI-95X52-56, SI-95X58-62, and SI-95X64-68. Their EGFR binding domains are derived from a humanized de-matured cetuximab variant (hDC-EGFR), namely SI-95m4-10. SI-95X45-49 has an anti-ABT-806 scFv on the antibody heavy chain fused to the C-terminus of anti-hDC-EGFR at the Fab position, SI-95X52-56 contains an anti-hDC-EGFR scFv on the antibody heavy chain fused to the N-terminus of ABT-806V9 at the Fab position, SI-95X58-62 contains an ABT-806V9 scFv on the antibody light chain fused to the C-terminus of anti-hDC-EGFR at the Fab position, and SI-95X64-68 contains an anti-hDC-EGFR scFv on the antibody heavy chain fused to the C-terminus of ABT-806 at the Fab position. The materials and methods used to generate the anti-EGFR×EGFRvIII antibodies are the same as or similar to those described in Example 1.

[0176] After transient expression in ExpiCHO cells, the titers were quantified using biolayer interferometry (Table 17). As expected, these bispecific antibodies (SI-95X40-68) were produced with variable efficiencies, which mainly depended on the composition and configuration of each binding domain. For example, molecules with an anti-hDC-EGFR scFv fused to the C-terminus of the antibody heavy chain (SI-95X63-68) appeared to be expressed less than molecules with an anti-hDC-EGFR scFv fused to the N-terminus of the heavy chain (SI-95X51-56). Additionally, molecules with an anti-hDC-EGFR Y101A domain (SI-95X47, 53, 58, 65) tended to have high titers, while molecules with an anti-hDC-EGFR Y104L domain (SI-95X46, 56, 61, 68) tended to have low titers.

[0177] After the first step of Protein A purification, the stability and aggregation of the anti-EGFR×EGFRvIII bispecific antibody were immediately evaluated by analytical size exclusion chromatography on a Waters UPLC system (Table 17). After the first step of purification, the conformation of the molecule seems to play the biggest role in the % of protein of interest. A large amount of high molecular weight aggregates demonstrated the instability of the SI-95X57-62 molecule. This trend demonstrated that the ABT-806V9 scFv fusion on the antibody light chain would produce generally unstable molecules. In contrast, SI-95X51-56 showed a very high % of protein of interest after the initial purification, indicating that the fusion of the anti-hDC-EGFR scFv to the N-terminus of the heavy chain of the antibody would generally produce stable molecules.

[0178] Biolayer interferometry was used to evaluate the affinity of the bispecific antibody for EGFR (Table 18). Molecules containing the anti-hDC-EGFR D103Y domain (SI-95X45, 55, 60, 67) showed no or very little binding to EGFR. This indicated that the affinity of this domain for EGFR was very low or eliminated. When compared to control molecules in the "matured" conformation, all other molecules containing the anti-hDC-EGFR domain showed a reduced affinity for EGFR. The reduction in affinity varied according to the conformation of the molecule and the type of anti-hDC-EGFR domain.

[0179] Biolayer interferometry was used to evaluate the affinity of bispecific antibodies for EGFRvIII (Table 19). Molecules containing the anti-hDC-EGFR D103Y domain (SI-95X45, 55, 60, 67) showed binding to EGFRvIII but not to EGFR (Table 18, Table 19). Collectively, this indicates that the ABT-806 domain (or domain derivative) is functional while anti-hDC-EGFR D103Y is non-functional or can function slightly depending on the molecular configuration. All molecules were able to bind EGFRvIII (although with slightly lower affinity) compared to the control molecules in the matured configuration. When compared to the monospecific mAbs in Table 14, binding was greatly enhanced in all bispecific antibodies. Thus, even with the anti-hDC-EGFR domain, these molecules can bind tightly to both EGFR and EGFRvIII. In fact, compared to wild-type cetuximab and ABT806 mAb, the affinity of some bispecific antibodies for EGFRvIII was enhanced, which replicated the multivalent avidity effect observed in Example 5 and demonstrated the mechanism by which matured cetuximab and ABT806 domains bind to the same EGFRvIII molecule simultaneously. Additionally, the binding of some antibodies to EGFRvIII (Table 19) was tighter than to EGFRwt (Table 18), indicating preferential binding to EGFRvIII while still maintaining good binding to EGFRwt.

[0180] Example 11: Dual-epitope tetravalent antibody exerts ADCC activity against EGFRvIII-expressing cells

[0181] To analyze any potency changes of the bispecific tetravalent antibodies when targeting tumor cells expressing EGFRwt and EGFRvIII, an ADCC assay was performed by co-culturing NK cells (1 donor) with U87-MG-GFP cells expressing EGFRwt, U87-MG-EGFRvIII-NR cells expressing both EGFRwt and EGFRvIII (i.e., mKATE2+), or a mixed culture of both. The mixed cell cultures were seeded and started with a 50:50 green / red signal. The green signal from U87-MG-GFP cells is related to live cells expressing EGFRwt, while the red signal from U87-MG-EGFRvIII-NR cells is related to cells co-expressing EGFRwt and EGFRvIII. After spheroid formation, NK cells were plated at an effector-to-target ratio of 5:1 and the antibodies were plated according to an 8-point log10 dose-response curve. After 60 hours, cell fluorescence was measured to quantify live cells and normalized relative to the 0-hour time point. This ADCC assay was used to screen monospecific bivalent antibodies (i.e., mAbs) as well as bispecific tetravalent antibodies as therapeutic candidates (Table 16).

[0182] The SI-95m4 and SI-95m5 antibodies are monospecific bivalent antibodies targeting EGFRvIII and EGFRwt, respectively. As Figure 4A shown, SI-95m4 could not kill U87-MG-EGFRwt-GFP cells in the ADCC assay, confirming that this humanized anti-EGFRvIII antibody retained the binding specificity unique to the ABT-806 antibody. However, when mixed with U87-MG-EGFRvIII-NR cells, SI-95m4 exerted ADCC activity against U87-MG-EGFRwt-GFP cells (i.e., EGFRwt target cells, Figure 4A ). This surprising result could be explained by the bystander effect of activated NK cells on EGFRwt-expressing cells. In contrast, SI-95m5 was able to kill both U87-MG-EGFRwt and U87-MG-EGFRvIII cells, either alone or in combination, which was consistent with the ability of this antibody to bind both EGFRwt and EGFRvIII ( Figure 4B ).

[0183] To analyze the effect of de-maturation, a panel of monospecific bivalent antibodies with mature EGFRwt (cetuximab) or de-matured EGFRwt binding domains were assayed for ADCC using U87-MG-GFP cells expressing EGFRwt and U87-MG-NR cells expressing EGFRwt / EGFRvIII, respectively. As shown in Figure 5, a panel of monospecific bivalent antibodies consisting of SI-95m6, SI-95m7, SI-95m8, SI-95m9, and SI-95m10 (Table 16) showed a series of reduced ADCC activities against EGFRwt-expressing cells when compared to the parental anti-hDC-EGFR antibody SI-95m5 ( Figure 5A ). Compared to the EC50 value of the parental mAb SI-95m5, the EC50 values of the variants increased up to 12-fold (for SI-95m6), indicating weaker potency. As expected, SI-95m4, an anti-EGFRvIII control antibody used in the assay, did not show activity. When EGFRwt / EGFRvIII-expressing cells were used, SI-95m6, SI-95m9, and SI-95m10 exerted their ADCC activities at levels similar to the positive control antibody SI-95m4 ( Figure 5B ). Among this panel of antibodies, SI-95m7 showed a right-shifted curve with a 3-fold increase in EC50, indicating a characteristic reduction in ADCC activity. Meanwhile, SI-95m8 no longer showed any activity against EGFR ( Figure 5A and Figure 5B), which may be due to a specific D103Y mutation in its EGFR binding domain. Even with limited data, when comparing a group of mAbs with different de-maturation mutations, there does not seem to be a correlated reduction in ADCC between EGFRwt expression and EGFRvIII expression. This result can be explained by the different densities or geometries of EGFRwt / EGFRvIII on the two cell populations, allowing for different binding affinities and initiation of ADCC.

[0184] To screen bispecific candidates, a group of bispecific tetravalent antibodies were constructed based on a group of monovalent epitope bivalent antibodies (Table 16, Figures 4 and 5), and the same ADCC assays were performed using EGFRwt-expressing cells and EGFRwt / EGFRvIII-expressing cells, respectively.

[0185] A group of bispecific tetravalent antibodies includes SI-95X52, SI-95X53, SI-95X54, SI-95X55, and SI-95X56, each of which contains an anti-EGFRvIII Fab binding domain, i.e., the hABT-806 (V9) antibody backbone, and an anti-EGFRwt scFv binding domain. This group shares a common antibody backbone derived from its parental mAb SI-94m4. Their EGFRwt binding domains are derived from an anti-hDC-EGFR scFv variant and correspond to a group of de-matured anti-EGFR wt mAbs, among which SI-95m5 is one of the parental antibodies, namely SI-95m10 (corresponding to SI-95X52), SI-95m6 (SI-95X53), SI-95m7 (SI-95X54), SI-95m8 (SI-95X55), and SI-95m9 (SI-95X56) (Table 16, Figure 5B ). At this point, SI-95X44 was used as a matured bispecific tetravalent antibody control (Table 16). These bispecific tetravalent antibodies showed a range of ADCC activities against EGFRwt-expressing cells ( Figure 6A ), and showed seemingly overlapping curves against tumor cells expressing EGFRvIII ( Figure 6B ). Notably, SI-95X55 had significantly reduced activity against EGFRwt-expressing cells. The EC50 values of these overlapping curves were calculated and listed in Figure 6BAmong them, EC50 values lower than the 19.3 nM EC50 of their parental antibody SI-95m4 were revealed. The lower EC50 values indicate improved ADCC activity. This observation may imply a synergistic effect of enhanced ADCC activity as a result of simultaneous binding to two EGFR epitopes on EGFRwt / EGFRvIII-expressing cells. Notably, in terms of the geometry and distance of the anti-EGFR and anti-EGFRvIII binding domains, different configurations of the bispecific antibody may allow the ability to alter this mechanism of simultaneous binding to cells, which may affect the degree of ADCC against tumor cells expressing EGFRwt as well as tumor cells expressing EGFRwt / EGFRvIII. Together with the findings in the mAb format, the synergistic effect of the bispecific tetravalent antibody may be affected by structural changes as a result of humanization, de-maturation, and / or configuration. Additionally, compared with SI-95m4 (19.39 nM) or SI-95X44 (14.72 nM) listed in Figure 6B SI-95X55 showed improved potency against EGFRwt / EGFRvIII-expressing cells, as shown by its EC50 (12.37 nM). This observation indicates that even the residual affinity / avidity for EGFRwt (Table 15) helps improve its ADCC potency. It also demonstrates that the therapeutic index was improved by reducing the potency against cells carrying EGFRwt while increasing the potency against cells carrying EGFRwt / EGFRvIII.

[0186] Example 12: Dual-epitope tetravalent antibody targeting tumors expressing both EGFRwt and EGFRvIII.

[0187] The results of Example 11 indicate that the ADCC activity of de-matured monospecific antibodies may not predict the same outcome for bispecific antibodies containing the same de-maturation mutations. For example, SI-95m8, SI-95X55, and SI-95X67 (with a configuration opposite to that of SI-95X55, see Table 16) contain the same D103Y mutation. The loss of ADCC potency against EGFRwt-expressing cells makes SI-95X55 a candidate for targeting only EGFRvIII tumors. In this case, the screening of bispecific tetravalent antibodies that can effectively target tumors expressing both EGFRwt and EGFRvIII focused on those antibody groups containing the de-maturation mutations Y101A and Y102A.

[0188] The Y101A mutation is a common feature of the de-matured antibodies SI-95m6 and SI-95X53 / SI-95X65, where the antibody backbone of SI-95X65 is not humanized (Table 16). As shown in Figure 7, SI-95X65 contains an anti-EGFR scFv at the other end of the heavy chain like SI-95X53, compared with SI-95X53 (Figure 7A ) showed overlapping but not identical killing curves compared to, and the EC50 of SI-95m6 against EGFRwt cells (199.9 nM) was almost 10-fold higher than that of the parental antibody SI-95m5 (EC50 = 16.06 nM) (i.e., mature binding to EGFRwt) (see Figure 7B the table in). This means that when the de-maturation mutation Y101A was placed in the bispecific tetravalent antibody, the EC50 (7.659 nM) was similar to that of the parental SI-95m5 antibody and the bispecific antibody (SI-95X44) containing the parental mAb backbone structure (EC50 = 14.38 nM). In this regard, SI-95X52 with the N92F mutation (Table 16) showed similar potency and complete killing ( Figure 6B ).

[0189] The Y102A mutation is a common feature of the de-maturation antibodies SI-95m7, SI-95X54, and SI-95X66, where the antibody backbone of SI-95X66 is not humanized and contains an anti-EGFR scFv domain at the other end of the heavy chain when compared to SI-95X54 (Table 16). When compared with the overlapping curves of SI-95m5 and SI-95X44, both SI-95m7 and SI-95X66 showed right-shifted curves with incomplete killing, while SI-95X54 had a complete killing curve for EGFRwt-expressing cells in the range of 10 nM to 100 nM (see Figure 7B the table in). The right shift of SI-95X54 was measured to be approximately 5-fold higher than the EC50 values of both SI-95M5 and SI-95X44. SI-95X54 is a bispecific tetravalent antibody whose scFv domain is derived from an anti-hDC-EFGR variant containing Y102A. Therefore, SI-95X54 revealed its EGFRvIII-biased cell killing while also killing EGFRwt tumors with an EC50 value at least 5-fold higher.

[0190] To further compare the changes in ADCC activity, a separate and mixed set of EGFRwt and EGFRvIII cells was used to evaluate the bispecific candidates. As a control, SI-95X44 showed overlapping curves and ED50 values similar to those expected ( Figure 8A ). Bispecific candidates such as SI-95X54 showed characteristic right-shifted curves and completely killed the mixed cells ( Figure 8B ). This result demonstrated that, as an exemplary bispecific tetravalent antibody, SI-95X54 was able to effectively kill a mixed population of EGFRwt and EGFRvIII cells, which could mimic the heterogeneity of glioblastoma in human patients, and the reduced sensitivity of killing EGFRwt cells might imply reduced toxicity to the healthy tissues of patients.

[0191] EGFRwt and EGFRvIII play complex and interacting roles in the development of brain tumors. In most cases of glioblastoma multiforme, EGFR overexpression is prevalent in tumor cells, while when stained with pan-EGFR antibodies, EGFRvIII usually shows patchy tumor positivity only in a few tumor cells. Such tumor heterogeneity remains an unresolved challenge in developing effective and meaningful treatments for brain tumors. Therefore, therapeutic antibodies that simultaneously target both EGFRwt and EGFRvIII cells and have EGFRvIII-biased binding may be an ideal solution. In this context, the present application provides a proof-of-concept for a therapeutic strategy by which bispecific tetravalent antibodies are generated to effectively eradicate mixed tumor cell populations expressing EGFRwt and / or EGFRvIII with low toxicity to normal tissues.

[0192] Table

[0193] Table 1. Configurations of bispecific tetravalent antibodies

[0194]

[0195]

[0196] Table 2. Stability assessment of bispecific tetravalent antibodies targeting EGFR-WT×EGFRvIII by expression titer and analytical size exclusion chromatography after the first-step protein A purification

[0197]

[0198]

[0199] Table 3. Binding kinetics of bispecific tetravalent antibodies to EGFRwt

[0200]

[0201]

[0202] Table 4. Binding kinetics of bispecific tetravalent antibodies to EGFRvIII

[0203]

[0204]

[0205] Table 5. EGFRwt epitope grouping of EGFRwt-targeting antibodies and EGFRvIII-targeting antibodies

[0206]

[0207] Table 6. EGFRvIII epitope grouping of EGFRwt-targeting antibodies and EGFRvIII-targeting antibodies

[0208]

[0209]

[0210] Table 7. Characterization of humanized ABT-806 variants, such as expression titer and purity evaluated by using analytical size exclusion chromatography after protein A purification

[0211]

[0212] Table 8. Binding affinity of humanized ABT-806 antibody to EGFRvIII

[0213]

[0214]

[0215] Table 9. Cell-binding assay confirmed that the antibody maintained binding to the antigen on the cell. All reported values were MFIs at 1.1 ug / mL for each antibody.

[0216] Humanized form EGFR EGFRvIII uPAR V1 634 10695 166 V2 661 11707 199 V3 1261 14652 204 V4 620 12031 195 V5 528 10224 190 V6 664 11803 185 V7 749 11179 196 V8 876 13080 196 V9 2713 16041 114 ABT-806 914 13986 184 Cetuximab 8896 9229 177

[0217] Table 10. Similarity of the lead humanized ABT-806 sequence to all sequences in the LENS database.

[0218]

[0219] Table 11. Energy calculation of the de-matured cetuximab mutations

[0220]

[0221]

[0222] Table 12. Characterization of the de-matured cetuximab variants, such as expression titer and purity evaluated by using analytical size exclusion chromatography after protein A purification

[0223]

[0224] Table 13. Binding affinity and avidity of de-matured cetuximab to EGFR

[0225]

[0226]

[0227] Table 14. Binding affinity and avidity of de-matured cetuximab to EGFRvIII

[0228]

[0229]

[0230] Table 15. Binding assay on cells confirms that the antibody maintains binding to the antigen on cells. All values reported are MFIs at 10 ug / mL for each antibody.

[0231]

[0232] Table 16. Configurations of anti-EGFR×EGFRvIII bispecific antibodies and component mAbs, which comprise a humanized de-matured EGFR-binding domain and a humanized EGFRvIII-binding domain

[0233]

[0234]

[0235]

[0236]

[0237] Table 17. Characterization of anti-EGFR×EGFRvIII bispecific antibodies comprising a humanized de-matured EGFR-binding domain and a humanized EGFRvIII-binding domain, such as expression titer and purity evaluated by using analytical size exclusion chromatography after protein A purification

[0238]

[0239]

[0240] Table 18. Binding kinetics of anti-EGFR×EGFRvIII bispecific tetravalent antibodies comprising a humanized de-matured EGFR-binding domain and a humanized EGFRvIII-binding domain to EGFR-WT

[0241]

[0242]

[0243] Table 19. Binding kinetics of anti-EGFR×EGFRvIII bispecific antibodies comprising a humanized de-matured EGFR-binding domain and a humanized EGFRvIII-binding domain to EGFRvIII

[0244]

[0245]

[0246] References

[0247] 1. Hanif F, Muzaffar K, Perveen K, Malhi SM, Simjee SU. Glioblastoma multiforme: A review of its epidemiology and pathogenesis through clinical presentation and treatment. Asian Pacific J Cancer Prev 2017; 18: 3–9.

[0248] 2. Gan HK, Cvrljevic AN, Johns TG. The epidermal growth factor receptor variant III (EGFRvIII): Where wild things are altered. FEBS J 2013; 280: 5350–70.

[0249] 3. Ramakrishnan MS, Eswaraiah A, Crombet T, Piedra P, Saurez G, Iyer H, Arvind AS. Nimotuzumab, a promising therapeutic monoclonal for treatment of tumors of epithelial origin. MAbs [Internet] 2009; 1: 41–8. Available from: www.landesbioscience.com

[0250] 4.AbbVie Provides Update on Depatuxizumab Mafodotin(Depatux-M),anInvestigational Medicinefor Newly Diagnosed Glioblastoma,an Aggressive Formof Brain Cancer[Internet].AbbVie PressRelease2019[cited 2022Mar 31];Availablefrom:https: / / news.abbvie.com / news / press-releases / abbvie-provides-update-on-depatuxizumab-mafodotin-depatux-m-an-investigational-medicine-for-newly-diagnosed-glioblastoma-an-aggressive-form-brain-cancer.htm

[0251] 5.Ellwanger K,Reusch U,Fucek I,Knackmuss S,Weichel M,Gantke T,Molkenthin V,ZhukovskyEA,Tesar M,Treder M.Highly specific and effectivetargeting of EGFRvIII-positive tumors withTandAb antibodies.Front Oncol 2017;7:1–17.

[0252] 6.Gedeon PC,Schaller TH,Chitneni SK,Choi BD,Kuan CT,Suryadevara CM,Snyder DJ,Schmittling RJ,Szafranski SE,Cui X,et al.A rationally designedfully human EGFRvIII:CD3-targeted bispecific antibody redirects human T cellsto treat patient-derived intracerebral malignantglioma.Clin Cancer Res 2018;24:3611–31.

[0253] 7. Li, S.; Schmitz, K. R.; Jeffrey, P. D.; Wiltzius, J. J. W.; Kussie, P.; Ferguson, K. M. Structural basis for inhibition of the epidermal growth factor receptor by Cetuximab. Cancer Cell 2005, 7, 301–311, doi:10.1016 / j.ccr.2005.03.003.

[0254] 11. Sickmier, E. A.; Kurzeja, R. J. M.; Michelsen, K.; Vazir, M.; Yang, E.; Tasker, A. S. The panitumumab EGFR complex reveals a binding mechanism that overcomes Cetuximab induced resistance. PLoS One 2016, 11, 1–11, doi:10.1371 / journal.pone.0163366.

[0255] 12. Bagchi, A.; Haidar, J. N.; Eastman, S. W.; Vieth, M.; Topper, M.; Iacolina, M. D.; Walker, J. M.; Forest, A.; Shen, Y.; Novosiadly, R. D.; et al. Molecular basis for necitumumab inhibition of EGFR variants associated with acquired Cetuximab resistance. Mol. Cancer Ther. 2018, 17, 521–531, doi:10.1158 / 1535-7163.MCT-17-0575.

[0256] Sequence Listing

[0257]

[0258]

[0259]

[0260]

[0261]

[0262] >Seq ID 1: Amino acid sequence of the heavy chain of cetuximab

[0263] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0264] >Seq ID 2: Nucleotide sequence of the heavy chain of cetuximab

[0265]

[0266] >Seq ID 3: Amino acid sequence of the light chain of cetuximab

[0267] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0268] >Seq ID 4: Nucleotide sequence of the light chain of cetuximab

[0269] GACATCTTGCTGACTCAGTCTCCAGTCATCCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGTATTGGCACAAACATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATTCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTGCAGATTATTACTGTCAACAAAATAATAACTGGCCAACCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0270] >Seq ID 5: Amino acid sequence of the heavy chain of nimotuzumab

[0271] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0272] >Seq ID 6: Nucleotide sequence of the heavy chain of nimotuzumab

[0273]

[0274] >Seq ID 7: Amino acid sequence of the light chain of nimotuzumab

[0275] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0276] >Seq ID 8: Nucleotide sequence of the light chain of nimotuzumab

[0277] GATATTCAAATGACTCAATCTCCTTCTTCTCTTTCTGCTTCTGTTGGTGATCGTGTTACTATTACTTGTCGTTCTTCTCAAAATATTGTTCATTCTAATGGTAATACTTATCTTGATTGGTATCAACAAACTCCTGGTAAAGCTCCTAAACTTCTTATTTATAAAGTTTCTAATCGTTTTTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTGGTTCTGGTACTGATTTTACTTTTACTATTTCTTCTCTTCAACCTGAAGATATTGCTACTTATTATTGTTTTCAATATTCTCATGTTCCTTGGACTTTTGGTCAAGGTACTAAACTTCAAATTACTCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0278] >Seq ID 9: Amino acid sequence of the heavy chain of ABT-806

[0279] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0280] >Seq ID 10: Nucleotide sequence of the heavy chain of ABT - 806

[0281]

[0282] >Seq ID 11: Amino acid sequence of the light chain of ABT-806

[0283] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0284] >Seq ID 12: Nucleotide sequence of the light chain of ABT-806

[0285] GACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG

[0286] >Seq ID 13: SI-95X1 heavy chain amino acid sequence

[0287] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0288] >Seq ID 14: SI-95X1 heavy chain nucleotide sequence

[0289]

[0290] >Seq ID 15: SI-95X2 heavy chain amino acid sequence

[0291] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0292] >Seq ID 16: SI-95X2 heavy chain nucleotide sequence

[0293]

[0294] >Seq ID 17: SI-95X3 heavy chain amino acid sequence

[0295] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK

[0296] >Seq ID 18: SI-95X3 heavy chain nucleotide sequence

[0297]

[0298] >Seq ID 19: SI-95X4 Heavy Chain Amino Acid Sequence

[0299] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQIT

[0300] >Seq ID 20: SI-95X4 Heavy Chain Nucleotide Sequence

[0301]

[0302] >Seq ID 21: Amino acid sequence of the heavy chain of SI-95X5

[0303] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0304] >Seq ID 22: Nucleotide sequence of the heavy chain of SI-95X5

[0305]

[0306] >Seq ID 23: SI-95X6 Heavy Chain Amino Acid Sequence

[0307] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0308] >Seq ID 24: SI-95X6 Heavy Chain Nucleotide Sequence

[0309]

[0310] >Seq ID 25: SI-95X7 Heavy Chain Amino Acid Sequence

[0311] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0312] >Seq ID 26: SI-95X7 Heavy Chain Nucleotide Sequence

[0313]

[0314] >Seq ID 27: Amino acid sequence of the heavy chain of SI-95X8

[0315] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0316] >Seq ID 28: Nucleotide sequence of the heavy chain of SI-95X8

[0317]

[0318] >Seq ID 29: SI-95X9 Heavy Chain Amino Acid Sequence

[0319] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0320] >Seq ID 30: SI-95X9 Heavy Chain Nucleotide Sequence

[0321]

[0322] >Seq ID 31: SI-95X10 Heavy Chain Amino Acid Sequence

[0323] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0324] >Seq ID 32: SI-95X10 Heavy Chain Nucleotide Sequence

[0325]

[0326] >Seq ID 33: Amino acid sequence of the light chain of SI-95X11

[0327] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0328] >Seq ID 34: Nucleotide sequence of the light chain of SI-95X11

[0329]

[0330] >Seq ID 35: Amino acid sequence of the light chain of SI-95X12

[0331] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0332] >Seq ID 36: Nucleotide sequence of the light chain of SI-95X12

[0333]

[0334] >Seq ID 37: Amino acid sequence of the light chain of SI-95X13

[0335] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0336] >Seq ID 38: Nucleotide sequence of the light chain of SI-95X13

[0337]

[0338] >Seq ID 39: Amino acid sequence of the light chain of SI-95X14

[0339] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0340] >Seq ID 40: Nucleotide sequence of the light chain of SI-95X14

[0341]

[0342] >Seq ID 41: Amino acid sequence of the light chain of SI-95X15

[0343] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0344] >Seq ID 42: Nucleotide sequence of the light chain of SI-95X15

[0345]

[0346] >Seq ID 43: Amino acid sequence of the light chain of SI-95X16

[0347] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0348] >Seq ID 44: Nucleotide sequence of the light chain of SI-95X16

[0349]

[0350] >Seq ID 45: Amino acid sequence of the heavy chain of SI-95X17

[0351] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKSGSGSGGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0352] >Seq ID 46: Nucleotide sequence of the heavy chain of SI-95X17

[0353]

[0354] >Seq ID 47: Amino acid sequence of the heavy chain of SI-95X18

[0355] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGSGSGGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0356] >Seq ID 48: Nucleotide sequence of the heavy chain of SI-95X18

[0357]

[0358] >Seq ID 49: Amino acid sequence of the heavy chain of SI-95X19

[0359] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0360] >Seq ID 50: Nucleotide sequence of the heavy chain of SI-95X19

[0361]

[0362] >Seq ID 51: Amino acid sequence of the heavy chain of SI-95X20

[0363] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSASGSGSGGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0364] >Seq ID 52: Nucleotide sequence of the heavy chain of SI-95X20

[0365]

[0366] >Seq ID 53: Amino acid sequence of the heavy chain of SI-95X21

[0367] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGSGSGGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0368] >Seq ID 54: Nucleotide sequence of the heavy chain of SI-95X21

[0369]

[0370] >Seq ID 55: SI-95X22 heavy chain amino acid sequence

[0371] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSQVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0372] >Seq ID 56: SI-95X22 heavy chain nucleotide sequence

[0373]

[0374] >Seq ID 57: Amino acid sequence of the light chain of SI-95X23

[0375] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKSGSGSGGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0376] >Seq ID 58: Nucleotide sequence of the light chain of SI-95X23

[0377]

[0378] >Seq ID 59: Amino acid sequence of the light chain of SI-95X24

[0379] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGSGSGGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0380] >Seq ID 60: Nucleotide sequence of the light chain of SI-95X24

[0381]

[0382] >Seq ID 61: Amino acid sequence of the light chain of SI-95X25

[0383] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0384] >Seq ID 62: Nucleotide sequence of the light chain of SI-95X25

[0385]

[0386] >Seq ID 63: Amino acid sequence of the light chain of SI-95X26

[0387] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSASGSGSGGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0388] >Seq ID 64: Nucleotide sequence of the light chain of SI-95X26

[0389]

[0390] >Seq ID 65: Amino acid sequence of the light chain of SI-95X27

[0391] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGSGSGGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0392] >Seq ID 66: Nucleotide sequence of the light chain of SI-95X27

[0393]

[0394] >Seq ID 67: Amino acid sequence of the light chain of SI-95X28

[0395] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0396] >Seq ID 68: Nucleotide sequence of the light chain of SI-95X28

[0397]

[0398] >Seq ID 69: Amino acid sequence of the light chain of SI-95X29

[0399] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0400] >Seq ID 70: Nucleotide sequence of the light chain of SI-95X29

[0401]

[0402] >Seq ID 71: Amino acid sequence of the light chain of SI-95X30

[0403] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0404] >Seq ID 72: Nucleotide sequence of the light chain of SI-95X30

[0405]

[0406] >Seq ID 73: Amino acid sequence of the light chain of SI-95X31

[0407] DIQMTQSPSSLSASVGDRVTITCRSSQNIVHSNGNTYLDWYQQTPGKAPKLLIYKVSNRFSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCFQYSHVPWTFGQGTKLQITRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0408] >Seq ID 74: Nucleotide sequence of the light chain of SI-95X31

[0409]

[0410] >Seq ID 75: SI-95X35 heavy chain amino acid sequence

[0411] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKGGGGSGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0412] >Seq ID 76: SI-95X35 heavy chain nucleotide sequence

[0413]

[0414] >Seq ID 77: Amino acid sequence of the light chain of SI-95X36

[0415] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKGGGGSGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSSGSGSGGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0416] >Seq ID 78: Nucleotide sequence of the light chain of SI-95X36

[0417]

[0418] >Seq ID 79: Amino acid sequence of the heavy chain of SI-95X37

[0419] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSASGSGSGGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0420] >Seq ID 80: Nucleotide sequence of the heavy chain of SI-95X37

[0421]

[0422] >Seq ID 81: Amino acid sequence of the light chain of SI-95X38

[0423] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSASGSGSGGGGGSGGGGSGGGGSDILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0424] >Seq ID 82: Nucleotide sequence of the light chain of SI-95X38

[0425]

[0426] >Seq ID 83: Amino acid sequence of the light chain of SI-95X39

[0427] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECSGSGSGGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0428] >Seq ID 84: Nucleotide sequence of the light chain of SI-95X39

[0429]

[0430] >Seq ID 85: SI-95X40 Heavy Chain Amino Acid Sequence

[0431] QVQLQQSGAEVKKPGSSVKVSCKASGYTFTNYYIYWVRQAPGQGLEWIGGINPTSGGSNFNEKFKTRVTITADESSTTAYMELSSLRSEDTAFYFCTRQGLWFDSDGRGFDFWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK

[0432] >Seq ID 86: SI-95X40 Heavy Chain Nucleotide Sequence

[0433]

[0434] >Seq ID 87: Amino acid sequence of the heavy chain of SI-95X41

[0435] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0436] >Seq ID 88: Nucleotide sequence of the heavy chain of SI-95X41

[0437]

[0438] >Seq ID 89: Amino acid sequence of the heavy chain of SI-95X42

[0439] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK

[0440] >Seq ID 90: Nucleotide sequence of the heavy chain of SI-95X42

[0441]

[0442] >Seq ID 91: Amino acid sequence of the light chain of SI-95X43

[0443] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELK

[0444] >Seq ID 92: Nucleotide sequence of the light chain of SI-95X43

[0445]

[0446] >Seq ID 93: SI-95X44 heavy chain amino acid sequence

[0447] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKGGGGSGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0448] >Seq ID 94: SI-95X44 heavy chain nucleotide sequence

[0449]

[0450] >Seq ID 95: Amino acid sequence of the light chain of SI-95X44

[0451] DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0452] >Seq ID 96: Nucleotide sequence of the light chain of SI-95X44

[0453] GACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0454] >Seq ID 97: SI-95X45 heavy chain amino acid sequence

[0455] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYYYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0456] >Seq ID 98: SI-95X45 heavy chain nucleotide sequence

[0457]

[0458] >Seq ID 99: Amino acid sequence of the light chain of SI-95m5

[0459] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0460] >Seq ID 100: Nucleotide sequence of the light chain of SI-95m5

[0461] GAGATCGTGCTGACCCAGTCTCCTTCCACACTGTCTGTGTCTCCCGGCGAGAGAGCCACCTTCAGCTGTAGAGCCTCTCAGTCCATCGGCACCAACATCCACTGGTATCAGCAGAAGCCCGGCAAGCCTCCTCGGCTGCTGATTAAGTACGCCTCCGAGTCCATCAGCGGCATCCCTGACAGATTCTCCGGCTCTGGCTCTGGCACCGAGTTTACCCTGACCATCTCCTCCGTGCAGTCCGAGGATTTCGCCGTGTACTACTGCCAGCAGAACAACAACTGGCCCACCACCTTTGGACCCGGCACCAAGCTGACCGTGCTGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0462] >Seq ID 101: SI-95X46 heavy chain amino acid sequence

[0463] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDLEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0464] >Seq ID 102: SI-95X46 heavy chain nucleotide sequence

[0465]

[0466] >Seq ID 103: Amino acid sequence of the heavy chain of SI-95X47

[0467] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTAYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0468] >Seq ID 104: Nucleotide sequence of the heavy chain of SI-95X47

[0469]

[0470] >Seq ID 105: SI-95X48 heavy chain amino acid sequence

[0471] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYADYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0472] >Seq ID 106: SI-95X48 heavy chain nucleotide sequence

[0473]

[0474] >Seq ID 107: Amino acid sequence of the heavy chain of SI-95X49

[0475] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAGGGGSGGGGSGGGGSGGGGSDILMTQSPSSMSVSLGDTVSITCHSSQDINSNIGWLQQRPGKSFKGLIYHGTNLDDEVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWTFGGGTKLEIK

[0476] >Seq ID 108: Nucleotide sequence of the heavy chain of SI-95X49

[0477]

[0478] >Seq ID 109: Amino acid sequence of the light chain of SI-95X49

[0479] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNFNWPTTFGPGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0480] >Seq ID 110: Nucleotide sequence of the light chain of SI-95X49

[0481] GAGATCGTGCTGACCCAGTCTCCTTCCACACTGTCTGTGTCTCCCGGCGAGAGAGCCACCTTCAGCTGTAGAGCCTCTCAGTCCATCGGCACCAACATCCACTGGTATCAGCAGAAGCCCGGCAAGCCTCCTCGGCTGCTGATTAAGTACGCCTCCGAGTCCATCAGCGGCATCCCTGACAGATTCTCCGGCTCTGGCTCTGGCACCGAGTTTACCCTGACCATCTCCTCCGTGCAGTCCGAGGATTTCGCCGTGTACTACTGCCAGCAGAACTTCAACTGGCCCACCACCTTTGGACCCGGCACCAAGCTGACCGTGCTGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0482] >Seq ID 111: SI-95m4 light chain amino acid sequence

[0483] DIVMTQSPSSVSLSLGDRVTITCHSSQDINSNIGWLQQKPGKAFKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLQAEDFATYYCVQYAQFPWTFGPGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0484] >Seq ID 112: SI-95m4 light chain nucleotide sequence

[0485] GACATCGTGATGACCCAATCTCCAAGCTCAGTGAGTCTGTCACTAGGCGACCGTGTCACCATTACATGTCATAGTTCCCAGGATATCAACAGTAATATTGGGTGGCTTCAGCAAAAGCCTGGTAAAGCCTTTAAAGGACTGATTTATCACGGGACCAACTTGGACGATGGAGTTCCCTCCCGGTTTTCTGGCTCCGGGTCCGGTGCAGACTATACTTTGACAATTAGCTCTCTCCAGGCAGAGGACTTCGCTACATATTACTGCGTCCAGTATGCCCAGTTCCCTTGGACCTTTGGACCCGGCACTAAACTGGAAATTAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0486] >Seq ID 113: Amino acid sequence of the heavy chain of SI-95X52

[0487] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNFNWPTTFGPGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0488] >Seq ID 114: SI-95X52 heavy chain nucleotide sequence

[0489]

[0490] >Seq ID 115: Amino acid sequence of the heavy chain of SI-95X53

[0491] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTAYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0492] >Seq ID 116: Nucleotide sequence of the heavy chain of SI-95X53

[0493]

[0494] >Seq ID 117: Amino acid sequence of the heavy chain of SI-95X54

[0495] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYADYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0496] >Seq ID 118: Nucleotide sequence of the heavy chain of SI-95X54

[0497]

[0498] >Seq ID 119: Amino acid sequence of the heavy chain of SI-95X55

[0499] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYYYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0500] >Seq ID 120: Nucleotide sequence of the heavy chain of SI-95X55

[0501]

[0502] >Seq ID 121: Amino acid sequence of the heavy chain of SI-95X56

[0503] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDLEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0504] >Seq ID 122: Nucleotide sequence of the heavy chain of SI-95X56

[0505]

[0506] >Seq ID 123: SI-95m5 heavy chain amino acid sequence

[0507] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0508] >Seq ID 124: SI-95m5 heavy chain nucleotide sequence

[0509]

[0510] >Seq ID 125: Amino acid sequence of the light chain of SI-95X57

[0511] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSGGGGSGGGGSGGGGSDIVMTQSPSSVSLSLGDRVTITCHSSQDINSNIGWLQQKPGKAFKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLQAEDFATYYCVQYAQFPWTFGPGTKLEIK

[0512] >Seq ID 126: Nucleotide sequence of the light chain of SI-95X50

[0513]

[0514] >Seq ID 127: Amino acid sequence of the heavy chain of SI-95X58

[0515] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTAYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0516] >Seq ID 128: Nucleotide sequence of the heavy chain of SI-95X58

[0517]

[0518] >Seq ID 129: SI-95X59 heavy chain amino acid sequence

[0519] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYADYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0520] >Seq ID 130: SI-95X59 heavy chain nucleotide sequence

[0521]

[0522] >Seq ID 131: SI-95X60 heavy chain amino acid sequence

[0523] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYYYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0524] >Seq ID 132: SI-95X60 heavy chain nucleotide sequence

[0525]

[0526] >Seq ID 133: SI-95X61 Heavy Chain Amino Acid Sequence

[0527] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDLEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0528] >Seq ID 134: SI-95X61 Heavy Chain Nucleotide Sequence

[0529]

[0530] >Seq ID 135: Amino acid sequence of the light chain of SI-95X62

[0531] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNFNWPTTFGPGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSGGGGSGGGGSGGGGSDIVMTQSPSSVSLSLGDRVTITCHSSQDINSNIGWLQQKPGKAFKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLQAEDFATYYCVQYAQFPWTFGPGTKLEIK

[0532] >Seq ID 136: Nucleotide sequence of the light chain of SI-95X62

[0533]

[0534] >Seq ID 137: Amino acid sequence of the heavy chain of SI-95X63

[0535] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVL

[0536] >Seq ID 138: Nucleotide sequence of the heavy chain of SI-95X63

[0537]

[0538] >Seq ID 139: Amino acid sequence of the heavy chain of SI-95X64

[0539] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNFNWPTTFGPGTKLTVL

[0540] >Seq ID 140: Nucleotide sequence of the heavy chain of SI-95X64

[0541]

[0542] >Seq ID 141: Amino acid sequence of the heavy chain of SI-95X65

[0543] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTAYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVL

[0544] >Seq ID 142: Nucleotide sequence of the heavy chain of SI-95X65

[0545]

[0546] >Seq ID 143: Amino acid sequence of the heavy chain of SI-95X66

[0547] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYADYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVL

[0548] >Seq ID 144: Nucleotide sequence of the heavy chain of SI-95X66

[0549]

[0550] >Seq ID 145: Amino acid sequence of the heavy chain of SI-95X67

[0551] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYYYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVL

[0552] >Seq ID 146: Nucleotide sequence of the heavy chain of SI-95X67

[0553]

[0554] >Seq ID 147: Amino acid sequence of the heavy chain of SI-95X68

[0555] DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWNWIRQFPGNKLEWMGYISYSGNTRYNPSLKSRISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDLEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSEIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVL

[0556] >Seq ID 148: Nucleotide sequence of the heavy chain of SI-95X68

[0557]

[0558] >Seq ID 149: Amino acid sequence of the heavy chain of SI-95m4

[0559] QVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0560] >Seq ID 150: Nucleotide sequence of the heavy chain of SI-95m4

[0561]

[0562] >Seq ID 151: Amino acid sequence of the light chain of SI-95m11

[0563] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNFNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0564] >Seq ID 152: Nucleotide sequence of the light chain of SI-95m11

[0565] GACATCTTGCTGACTCAGTCTCCAGTCATCCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGTATTGGCACAAACATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATTCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTGCAGATTATTACTGTCAACAAAATTTCAACTGGCCAACCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0566] >Seq ID 153: SI-95m12 light chain amino acid sequence

[0567] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNKNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0568] >Seq ID 154: SI-95m12 light chain nucleotide sequence

[0569] GACATCTTGCTGACTCAGTCTCCAGTCATCCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGTATTGGCACAAACATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATTCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTGCAGATTATTACTGTCAACAAAATAAGAACTGGCCAACCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0570] >Seq ID 155: Amino acid sequence of the heavy chain of SI-95m13

[0571] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTAYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0572] >Seq ID 156: SI-95m13 heavy chain nucleotide sequence

[0573]

[0574] >Seq ID 157: Amino acid sequence of the heavy chain of SI-95m14

[0575] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTWYDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0576] >Seq ID 158: Nucleotide sequence of the heavy chain of SI-95m14

[0577]

[0578] >Seq ID 159: Amino acid sequence of the heavy chain of SI-95m15

[0579] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYADYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0580] >Seq ID 160: Nucleotide sequence of the heavy chain of SI-95m15

[0581]

[0582] >Seq ID 161: Amino acid sequence of the heavy chain of SI-95m16

[0583] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYVDYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0584] >Seq ID 162: Nucleotide sequence of the heavy chain of SI-95m16

[0585]

[0586] >Seq ID 163: Amino acid sequence of the heavy chain of SI-95m17

[0587] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYFYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0588] >Seq ID 164: Nucleotide sequence of the heavy chain of SI-95m17

[0589]

[0590] >Seq ID 165: Amino acid sequence of the heavy chain of SI-95m18

[0591] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYWYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0592] >Seq ID 166: Nucleotide sequence of the heavy chain of SI-95m18

[0593]

[0594] >Seq ID 167: Amino acid sequence of the heavy chain of SI-95m19

[0595] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYYYEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0596] >Seq ID 168: Nucleotide sequence of the heavy chain of SI-95m19

[0597]

[0598] >Seq ID 169: Amino acid sequence of the heavy chain of SI-95m20

[0599] QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDLEFAYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0600] >Seq ID 170: Nucleotide sequence of the heavy chain of SI-95m20

[0601]

[0602] >Seq ID 171: Amino acid sequence of ABT-806 V1 heavy chain

[0603] QVQLQESGPGLVKPSQSLSLTCTVSGYSITSDFAWNWIRQPPGKGLEWIGYISYSGNTRYNPSLKSRVTISRDTSKNQFSLQLNSVTAEDTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0604] >Seq ID 172: Nucleotide sequence of ABT-806 V1 heavy chain

[0605]

[0606] >Seq ID 173: Amino acid sequence of the light chain of ABT-806V1

[0607] DIQMTQSPSSVSASVGDRVTITCHSSQDINSNIGWLQQKPGKAPKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLQPEDFATYYCVQYAQFPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0608] >Seq ID 174: Nucleotide sequence of the light chain of ABT-806V1

[0609] GACATTCAGATGACACAGAGCCCAAGTAGCGTTTCTGCCAGCGTGGGGGACCGAGTCACCATTACTTGTCACAGTTCACAAGATATTAACTCTAACATCGGCTGGCTGCAGCAAAAACCAGGTAAAGCACCAAAAGGGCTCATTTATCATGGGACCAACCTGGATGACGGAGTCCCGAGTCGATTCAGCGGGAGTGGAAGCGGAGCTGATTATACCCTGACGATCTCAAGCTTGCAGCCAGAGGATTTCGCCACATACTATTGTGTCCAGTATGCCCAGTTCCCCTGGACATTTGGCCAGGGTACGAAGGTGGAGATTAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0610] >Seq ID 175: Amino acid sequence of ABT-806V2 light chain

[0611] DIVMTQSPSSLSVSPGDRVTITCHSSQDINSNIGWLQQKPGKAPKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLEPEDFAVYYCVQYAQFPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0612] >Seq ID 176: Nucleotide sequence of ABT-806V2 light chain

[0613] GACATAGTGATGACACAGTCTCCAAGTTCTTTGAGCGTATCCCCAGGCGACCGCGTAACAATTACCTGTCACAGCTCCCAGGATATTAATAGTAATATTGGCTGGCTTCAACAGAAGCCAGGGAAGGCACCAAAAGGACTGATCTACCACGGAACTAATCTGGACGATGGGGTTCCTTCACGGTTCTCCGGGTCCGGCTCTGGTGCCGACTACACTCTGACAATTAGCTCCCTGGAGCCCGAGGACTTCGCAGTGTACTATTGCGTGCAGTACGCACAATTTCCCTGGACATTTGGTCAAGGCACCAAAGTCGAAATTAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0614] >Seq ID 177: Amino acid sequence of ABT-806 V4 heavy chain

[0615] QVQLQESGPGLVKPSQSLSLTCTVSGYSITSDFAWNWIRQAPGKGLEWVGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLQLNSVTAEDTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0616] >Seq ID 178: ABT-806V4 heavy chain nucleotide sequence

[0617]

[0618] >Seq ID 179: Amino acid sequence of the heavy chain of SI-95X51

[0619] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVLGGGGSGGGGSGGGGSGGGGSQVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0620] >Seq ID 180: Nucleotide sequence of the heavy chain of SI-95X51

[0621]

[0622] >Seq ID 201: Cetuximab VH Amino Acid Sequence

[0623] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYYDYEFAY WGQGTLVTVSS

[0624] >Seq ID 202: Cetuximab VH Nucleotide Sequence

[0625] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACTATGATTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0626] >Seq ID 203: Cetuximab VL Amino Acid Sequence

[0627] DILLTQSPVILSVSPGERVSFSC RASQSIGTNIH WYQQRTNGSPRLLIK YASESIS GIPSRFSGSGSGTDFTLSINSVESEDIADYYC QQNNNWPTT FGAGTKLELK

[0628] >Seq ID 204: Cetuximab VL Nucleotide Sequence

[0629] GACATCTTGCTGACTCAGTCTCCAGTCATCCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGTATTGGCACAAACATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATTCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTGCAGATTATTACTGTCAACAAAATAATAACTGGCCAACCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0630] >Seq ID 205: VH amino acid sequence of nimotuzumab

[0631] QVQLQQSGAEVKKPGSSVKVSCKASGYTFT NYYIY WVRQAPGQGLEWIG GINPTSGGSNFNEKFKT RVTITADESSTTAYMELSSLRSEDTAFYFCTR QGLWFDSDGRGFDF WGQGTTVTVSS

[0632] >Seq ID 206: VH nucleotide sequence of nimotuzumab

[0633] CAGGTGCAGCTGCAGCAGAGCGGCGCCGAGGTGAAGAAGCCCGGCAGCAGCGTGAAGGTGAGCTGCAAGGCCAGCGGCTACACCTTCACCAACTACTACATCTACTGGGTGCGGCAGGCCCCCGGCCAGGGCCTGGAGTGGATCGGCGGCATCAACCCCACCAGCGGCGGCAGCAACTTCAACGAGAAGTTCAAGACCCGGGTGACCATCACCGCCGACGAGAGCAGCACCACCGCCTACATGGAGCTGAGCAGCCTGCGGAGCGAGGACACCGCCTTCTACTTCTGCACCCGGCAGGGCCTGTGGTTCGACAGCGACGGCCGGGGCTTCGACTTCTGGGGCCAGGGCACCACCGTGACCGTGAGCAGC

[0634] >Seq ID 207: Amino acid sequence of the VL of nimotuzumab

[0635] DIQMTQSPSSLSASVGDRVTITC RSSQNIVHSNGNTYLD WYQQTPGKAPKLLIY KVSNRFS GVPSRFSGSGSGTDFTFTISSLQPEDIATYYC FQYSHVPWT FGQGTKLQIT

[0636] >Seq ID 208: Nucleotide sequence of the VL of nimotuzumab

[0637] GATATTCAAATGACTCAATCTCCTTCTTCTCTTTCTGCTTCTGTTGGTGATCGTGTTACTATTACTTGTCGTTCTTCTCAAAATATTGTTCATTCTAATGGTAATACTTATCTTGATTGGTATCAACAAACTCCTGGTAAAGCTCCTAAACTTCTTATTTATAAAGTTTCTAATCGTTTTTCTGGTGTTCCTTCTCGTTTTTCTGGTTCTGGTTCTGGTACTGATTTTACTTTTACTATTTCTTCTCTTCAACCTGAAGATATTGCTACTTATTATTGTTTTCAATATTCTCATGTTCCTTGGACTTTTGGTCAAGGTACTAAACTTCAAATTACT

[0638] >Seq ID 209: Amino acid sequence of the VH of ABT-806

[0639] DVQLQESGPSLVKPSQSLSLTCTVTGYSIT SDFAWN WIRQFPGNKLEWMG YISYSGNTRYNPSLKS RISITRDTSKNQFFLQLNSVTIEDTATYYCVT AGRGFPY WGQGTLVTVSA

[0640] >Seq ID 210: Nucleotide sequence of the VH of ABT-806

[0641] GATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA

[0642] >Seq ID 211: Amino acid sequence of ABT-806VL

[0643] DILMTQSPSSMSVSLGDTVSITC HSSQDINSNIG WLQQRPGKSFKGLIY HGTNLDD EVPSRFSGSGSGADYSLTISSLESEDFADYYC VQYAQFPWT FGGGTKLEIK

[0644] >Seq ID 212: Nucleotide sequence of ABT-806VL

[0645] GACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA

[0646] >Seq ID 213: Amino acid sequence of (G4S)2 linker

[0647] GGGGSGGGGS

[0648] >Seq ID 214: (G4S)2 linker nucleotide sequence

[0649] GGCGGTGGAGGGTCCGGCGGTGGTGGATCC

[0650] >Seq ID 215: (G4S)4 linker amino acid sequence

[0651] GGGGSGGGGSGGGGS

[0652] >Seq ID 216: (G4S)4 linker nucleotide sequence

[0653] GGTGGAGGAGGCAGTGGTGGTGGAGGAAGCGGAGGTGGTGGCAGC

[0654] >Seq ID 217: (G4S)5 linker amino acid sequence

[0655] GGGGSGSGSGGGGGSGGGGSGGGGS

[0656] >Seq ID 218: (G4S)5 linker nucleotide sequence

[0657] GGTGGCGGTGGCTCCGGATCCGGTTCTGGAGGAGGCGGTGGAAGCGGAGGCGGTGGCTCTGGAGGAGGCGGTTCG

[0658] >Seq ID 219: (G4S)6 linker amino acid sequence

[0659] GGGGSGGGGSGSGSGGGGGSGGGGSGGGGS

[0660] >Seq ID 220: (G4S)6 linker nucleotide sequence

[0661] GGAGGCGGTGGTAGCGGTGGCGGTGGCTCCGGATCCGGTTCTGGAGGAGGCGGTGGAAGCGGAGGCGGTGGCTCTGGAGGAGGCGGTTCG

[0662] >Seq ID 221: Human IgG1 amino acid sequence

[0663] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[0664] >Seq ID 222: Human IgG1 nucleotide sequence

[0665] GCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT

[0666] >Seq ID 223: Human CK amino acid sequence

[0667] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0668] >Seq ID 224: Human CK nucleotide sequence

[0669] CGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT

[0670] >Seq ID 225: ABT-806V1 VH amino acid sequence

[0671] QVQLQESGPGLVKPSQSLSLTCTVSGYSITSDFAWNWIRQPPGKGLEWIGYISYSGNTRYNPSLKSRVTISRDTSKNQFSLQLNSVTAEDTAVYYCVTAGRGFPYWGQGTLVTVSS

[0672] >Seq ID 226: ABT-806V1 VH nucleotide sequence

[0673] CAAGTGCAGTTGCAGGAGAGTGGTCCGGGCCTGGTGAAGCCTTCTCAGTCACTGAGCCTCACGTGCACCGTAAGCGGTTATAGTATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGCCGCCCGGGAAGGGACTGGAGTGGATCGGATACATTTCCTACAGTGGAAACACTCGATATAATCCTAGTCTTAAAAGTCGGGTAACCATCTCTAGGGATACCAGCAAGAATCAGTTCTCATTGCAACTCAATTCAGTGACCGCAGAAGACACCGCAGTCTACTATTGCGTCACAGCCGGACGCGGTTTCCCGTACTGGGGCCAAGGAACACTAGTCACCGTGTCTTCC

[0674] >Seq ID 227: Amino acid sequence of ABT-806V1 VL

[0675] DIQMTQSPSSVSASVGDRVTITCHSSQDINSNIGWLQQKPGKAPKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLQPEDFATYYCVQYAQFPWTFGQGTKVEIK

[0676] >Seq ID 228: Nucleotide sequence of ABT-806V1 VL

[0677] GACATTCAGATGACACAGAGCCCAAGTAGCGTTTCTGCCAGCGTGGGGGACCGAGTCACCATTACTTGTCACAGTTCACAAGATATTAACTCTAACATCGGCTGGCTGCAGCAAAAACCAGGTAAAGCACCAAAAGGGCTCATTTATCATGGGACCAACCTGGATGACGGAGTCCCGAGTCGATTCAGCGGGAGTGGAAGCGGAGCTGATTATACCCTGACGATCTCAAGCTTGCAGCCAGAGGATTTCGCCACATACTATTGTGTCCAGTATGCCCAGTTCCCCTGGACATTTGGCCAGGGTACGAAGGTGGAGATTAAG

[0678] >Seq ID 229: Amino acid sequence of ABT-806V2 VH

[0679] QVQLQESGPGLVKPSQSLSLTCTVSGYSITSDFAWNWIRQAPGKGLEWVGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLQLNSVTAEDTAVYYCVTAGRGFPYWGQGTLVTVSS

[0680] >Seq ID 230: ABT-806V2 VH nucleotide sequence

[0681] CAAGTCCAGCTGCAGGAAAGTGGCCCTGGACTCGTTAAGCCTAGTCAGAGCTTGTCATTGACTTGCACAGTCAGTGGTTATTCCATCACAAGTGACTTTGCCTGGAATTGGATTCGGCAGGCTCCCGGGAAGGGGCTGGAATGGGTCGGGTATATTTCCTACAGCGGAAATACTCGATATAACCCATCCCTTAAAAGTCGCTTCACCATATCTCGTGACACCAGCAAAAATCAATTTTATCTTCAACTCAATTCAGTGACAGCCGAGGATACAGCTGTTTACTATTGTGTGACAGCTGGCAGAGGATTTCCCTATTGGGGACAGGGAACCCTTGTAACGGTCTCCAGC

[0682] >Seq ID 231: ABT-806V2 VL amino acid sequence

[0683] DIVMTQSPSSLSVSPGDRVTITCHSSQDINSNIGWLQQKPGKAPKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLEPEDFAVYYCVQYAQFPWTFGQGTKVEIK

[0684] >Seq ID 232: ABT-806V2 VL nucleotide sequence

[0685] GACATAGTGATGACACAGTCTCCAAGTTCTTTGAGCGTATCCCCAGGCGACCGCGTAACAATTACCTGTCACAGCTCCCAGGATATTAATAGTAATATTGGCTGGCTTCAACAGAAGCCAGGGAAGGCACCAAAAGGACTGATCTACCACGGAACTAATCTGGACGATGGGGTTCCTTCACGGTTCTCCGGGTCCGGCTCTGGTGCCGACTACACTCTGACAATTAGCTCCCTGGAGCCCGAGGACTTCGCAGTGTACTATTGCGTGCAGTACGCACAATTTCCCTGGACATTTGGTCAAGGCACCAAAGTCGAAATTAAA

[0686] >Seq ID 233: ABT-806V3 VH amino acid sequence

[0687] QVQLQESGPSLVKPSQTLSLTCTVSGYSITSDFAWNWIRQFPGRKLEWMGYISYSGNTRYNPSLKSRFTISRDTSKNQFYLKLRSVTIADTAVYYCVTAGRGFPYWGQGTLVTVSS

[0688] >Seq ID 234: ABT-806V3 VH nucleotide sequence

[0689] CAAGTGCAGCTCCAGGAGTCAGGCCCTAGCCTCGTGAAGCCCTCCCAGACCCTGAGCCTGACCTGCACCGTATCTGGTTATAGTATTACAAGTGACTTCGCCTGGAACTGGATTAGGCAGTTTCCAGGTCGCAAGCTGGAGTGGATGGGGTATATCTCCTACTCCGGTAACACTCGATACAACCCTTCCCTCAAAAGCAGATTTACCATTTCCAGAGACACATCTAAGAACCAATTTTATCTGAAGTTGAGATCTGTTACCATCGCTGATACTGCTGTGTACTATTGCGTTACCGCCGGACGCGGCTTCCCCTATTGGGGGCAGGGAACCCTGGTGACTGTATCATCC

[0690] >Seq ID 235: Amino acid sequence of ABT-806V3 VL

[0691] DIVMTQSPSSVSLSLGDRVTITCHSSQDINSNIGWLQQKPGKAFKGLIYHGTNLDDGVPSRFSGSGSGADYTLTISSLQAEDFATYYCVQYAQFPWTFGPGTKLEIK

[0692] >Seq ID 236: Nucleotide sequence of ABT-806V3 VL

[0693] GACATCGTGATGACCCAATCTCCAAGCTCAGTGAGTCTGTCACTAGGCGACCGTGTCACCATTACATGTCATAGTTCCCAGGATATCAACAGTAATATTGGGTGGCTTCAGCAAAAGCCTGGTAAAGCCTTTAAAGGACTGATTTATCACGGGACCAACTTGGACGATGGAGTTCCCTCCCGGTTTTCTGGCTCCGGGTCCGGTGCAGACTATACTTTGACAATTAGCTCTCTCCAGGCAGAGGACTTCGCTACATATTACTGCGTCCAGTATGCCCAGTTCCCTTGGACCTTTGGACCCGGCACTAAACTGGAAATTAAA

[0694] >Seq ID 237: Amino acid sequence of SI-95m5 (humanized cetuximab) VH

[0695] QVQLQQSGPGLVKPSETLSITCTVSGFSLTNYGVHWIRQAPGKGLEWLGVIWSGGNTDYNTPFTSRFTITKDNSKNQVYFKLRSVRADDTAIYYCARALTYYDYEFAYWGQGTLVTVSS

[0696] >Seq ID 238: Nucleotide sequence of SI-95m5 (humanized cetuximab) VH

[0697] CAAGTTCAGTTGCAGCAGTCTGGCCCTGGCCTGGTCAAGCCTTCTGAGACACTGTCCATCACCTGTACCGTGTCCGGCTTCTCCCTGACCAATTACGGCGTGCACTGGATCAGACAGGCCCCTGGCAAAGGACTGGAATGGCTGGGAGTGATTTGGAGCGGCGGCAACACCGACTACAACACCCCTTTCACCAGCCGGTTCACCATCACCAAGGACAACTCCAAGAACCAGGTGTACTTCAAGCTGCGGAGCGTGCGGGCTGATGACACCGCCATCTACTACTGTGCTCGGGCCCTGACCTACTACGACTACGAGTTTGCTTACTGGGGCCAGGGCACCCTGGTCACAGTTTCTTCT

[0698] >Seq ID 239: SI-95m5 (Humanized Cetuximab) VL Amino Acid Sequence

[0699] EIVLTQSPSTLSVSPGERATFSCRASQSIGTNIHWYQQKPGKPPRLLIKYASESISGIPDRFSGSGSGTEFTLTISSVQSEDFAVYYCQQNNNWPTTFGPGTKLTVL

[0700] >Seq ID 240: SI-95m5 (Humanized Cetuximab) VL Nucleotide Sequence

[0701] GAGATCGTGCTGACCCAGTCTCCTTCCACACTGTCTGTGTCTCCCGGCGAGAGAGCCACCTTCAGCTGTAGAGCCTCTCAGTCCATCGGCACCAACATCCACTGGTATCAGCAGAAGCCCGGCAAGCCTCCTCGGCTGCTGATTAAGTACGCCTCCGAGTCCATCAGCGGCATCCCTGACAGATTCTCCGGCTCTGGCTCTGGCACCGAGTTTACCCTGACCATCTCCTCCGTGCAGTCCGAGGATTTCGCCGTGTACTACTGCCAGCAGAACAACAACTGGCCCACCACCTTTGGACCCGGCACCAAGCTGACCGTGCTG

[0702] >Seq ID 241: Cetuximab N92F VL Amino Acid Sequence

[0703] EIVLTQSPSTLSVSPGERATFSC RASQSIGTNIH WYQQKPGKPPRLLIK YASESIS GIPDRFSGSGSGTEFTLTISSVQSEDFAVYYC QQNFNWPTT FGPGTKLTVL

[0704] >Seq ID 242: Cetuximab N92F VL Nucleotide Sequence

[0705] GAGATCGTGCTGACCCAGTCTCCTTCCACACTGTCTGTGTCTCCCGGCGAGAGAGCCACCTTCAGCTGTAGAGCCTCTCAGTCCATCGGCACCAACATCCACTGGTATCAGCAGAAGCCCGGCAAGCCTCCTCGGCTGCTGATTAAGTACGCCTCCGAGTCCATCAGCGGCATCCCTGACAGATTCTCCGGCTCTGGCTCTGGCACCGAGTTTACCCTGACCATCTCCTCCGTGCAGTCCGAGGATTTCGCCGTGTACTACTGCCAGCAGAACTTCAACTGGCCCACCACCTTTGGACCCGGCACCAAGCTGACCGTGCTG

[0706] >Seq ID 243: Cetuximab N92K VL Amino Acid Sequence

[0707] DILLTQSPVILSVSPGERVSFSC RASQSIGTNIH WYQQRTNGSPRLLIK YASESIS GIPSRFSGSGSGTDFTLSINSVESEDIADYYC QQNKNWPTT FGAGTKLELK

[0708] >Seq ID 244: Cetuximab N92K VL Nucleotide Sequence

[0709] GACATCTTGCTGACTCAGTCTCCAGTCATCCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGTATTGGCACAAACATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATTCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTGCAGATTATTACTGTCAACAAAATAAGAACTGGCCAACCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

[0710] >Seq ID 245: Humanized cetuximab Y101A VH amino acid sequence

[0711] QVQLQQSGPGLVKPSETLSITCTVSGFSLT NYGVH WIRQAPGKGLEWLG VIWSGGNTDYNTPFTS RFTITKDNSKNQVYFKLRSVRADDTAIYYCAR ALTAYDYEFAY WGQGTLVTVSS

[0712] >Seq ID 246: Humanized cetuximab Y101A VH nucleotide sequence

[0713] CAAGTTCAGTTGCAGCAGTCTGGCCCTGGCCTGGTCAAGCCTTCTGAGACACTGTCCATCACCTGTACCGTGTCCGGCTTCTCCCTGACCAATTACGGCGTGCACTGGATCAGACAGGCCCCTGGCAAAGGACTGGAATGGCTGGGAGTGATTTGGAGCGGCGGCAACACCGACTACAACACCCCTTTCACCAGCCGGTTCACCATCACCAAGGACAACTCCAAGAACCAGGTGTACTTCAAGCTGCGGAGCGTGCGGGCTGATGACACCGCCATCTACTACTGTGCTCGGGCCCTGACCGCCTACGACTACGAGTTTGCTTACTGGGGCCAGGGCACCCTGGTCACAGTTTCTTCT

[0714] >Seq ID 247: Amino acid sequence of Cetuximab Y101W VH

[0715] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTWYDYEFAY WGQGTLVTVSS

[0716] >Seq ID 248: Nucleotide sequence of Cetuximab Y101W VH

[0717] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTGGTATGATTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0718] >Seq ID 249: Amino acid sequence of humanized Cetuximab Y102A VH

[0719] QVQLQQSGPGLVKPSETLSITCTVSGFSLT NYGVH WIRQAPGKGLEWLG VIWSGGNTDYNTPFTS RFTITKDNSKNQVYFKLRSVRADDTAIYYCAR ALTYADYEFAY WGQGTLVTVSS

[0720] >Seq ID 250: Nucleotide sequence of humanized Cetuximab Y102A VH

[0721] CAAGTTCAGTTGCAGCAGTCTGGCCCTGGCCTGGTCAAGCCTTCTGAGACACTGTCCATCACCTGTACCGTGTCCGGCTTCTCCCTGACCAATTACGGCGTGCACTGGATCAGACAGGCCCCTGGCAAAGGACTGGAATGGCTGGGAGTGATTTGGAGCGGCGGCAACACCGACTACAACACCCCTTTCACCAGCCGGTTCACCATCACCAAGGACAACTCCAAGAACCAGGTGTACTTCAAGCTGCGGAGCGTGCGGGCTGATGACACCGCCATCTACTACTGTGCTCGGGCCCTGACCTACGCCGACTACGAGTTTGCTTACTGGGGCCAGGGCACCCTGGTCACAGTTTCTTCT

[0722] >Seq ID 251: Cetuximab D103F VH Amino Acid Sequence

[0723] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYYFYEFAY WGQGTLVTVSS

[0724] >Seq ID 252: Cetuximab D103F VH Nucleotide Sequence

[0725] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACTATTTCTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0726] >Seq ID 253: Cetuximab D103W VH Amino Acid Sequence

[0727] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYYWYEFAY WGQGTLVTVSS

[0728] >Seq ID 254: Cetuximab D103W VH Nucleotide Sequence

[0729] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACTATTGGTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0730] >Seq ID 255: Humanized Cetuximab D103Y VH Amino Acid Sequence

[0731] QVQLQQSGPGLVKPSETLSITCTVSGFSLT NYGVH WIRQAPGKGLEWLG VIWSGGNTDYNTPFTS RFTITKDNSKNQVYFKLRSVRADDTAIYYCAR ALTYYYYEFAY WGQGTLVTVSS

[0732] >Seq ID 256: Humanized Cetuximab D103Y VH Nucleotide Sequence

[0733] CAAGTTCAGTTGCAGCAGTCTGGCCCTGGCCTGGTCAAGCCTTCTGAGACACTGTCCATCACCTGTACCGTGTCCGGCTTCTCCCTGACCAATTACGGCGTGCACTGGATCAGACAGGCCCCTGGCAAAGGACTGGAATGGCTGGGAGTGATTTGGAGCGGCGGCAACACCGACTACAACACCCCTTTCACCAGCCGGTTCACCATCACCAAGGACAACTCCAAGAACCAGGTGTACTTCAAGCTGCGGAGCGTGCGGGCTGATGACACCGCCATCTACTACTGTGCTCGGGCCCTGACCTACTACTACTACGAGTTTGCTTACTGGGGCCAGGGCACCCTGGTCACAGTTTCTTCT

[0734] >Seq ID 257: Humanized cetuximab Y104L VH amino acid sequence

[0735] QVQLQQSGPGLVKPSETLSITCTVSGFSLT NYGVH WIRQAPGKGLEWLG VIWSGGNTDYNTPFTS RFTITKDNSKNQVYFKLRSVRADDTAIYYCAR ALTYYDLEFAY WGQGTLVTVSS

[0736] >Seq ID 258: Humanized cetuximab Y104L VH nucleotide sequence

[0737] CAAGTTCAGTTGCAGCAGTCTGGCCCTGGCCTGGTCAAGCCTTCTGAGACACTGTCCATCACCTGTACCGTGTCCGGCTTCTCCCTGACCAATTACGGCGTGCACTGGATCAGACAGGCCCCTGGCAAAGGACTGGAATGGCTGGGAGTGATTTGGAGCGGCGGCAACACCGACTACAACACCCCTTTCACCAGCCGGTTCACCATCACCAAGGACAACTCCAAGAACCAGGTGTACTTCAAGCTGCGGAGCGTGCGGGCTGATGACACCGCCATCTACTACTGTGCTCGGGCCCTGACCTACTACGACCTCGAGTTTGCTTACTGGGGCCAGGGCACCCTGGTCACAGTTTCTTCT

[0738] >Seq ID 259: Cetuximab Y101A VH amino acid sequence

[0739] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTAYDYEFAY WGQGTLVTVSS

[0740] >Seq ID 260: Cetuximab Y101A VH nucleotide sequence

[0741] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCGCCTATGATTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0742] >Seq ID 261: Amino acid sequence of cetuximab Y102A VH

[0743] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYADYEFAY WGQGTLVTVSS

[0744] >Seq ID 262: Nucleotide sequence of cetuximab Y102A VH

[0745] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACGCCGATTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0746] >Seq ID 263: Cetuximab Y102V VH amino acid sequence

[0747] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYVDYEFAY WGQGTLVTVSS

[0748] >Seq ID 264: Cetuximab Y102V VH nucleotide sequence

[0749] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACGTGGATTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0750] >Seq ID 265: Cetuximab D103Y VH Amino Acid Sequence

[0751] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYYYYEFAY WGQGTLVTVSS

[0752] >Seq ID 266: Cetuximab D103Y VH Nucleotide Sequence

[0753] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACTATTACTACGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0754] >Seq ID 267: Cetuximab Y104L VH Amino Acid Sequence

[0755] QVQLKQSGPGLVQPSQSLSITCTVSGFSLT NYGVH WVRQSPGKGLEWLG VIWSGGNTDYNTPFTS RLSINKDNSKSQVFFKMNSLQSNDTAIYYCAR ALTYYDLEFAY WGQGTLVTVSS

[0756] >Seq ID 268: Cetuximab Y104L VH Nucleotide Sequence

[0757] CAGGTGCAGCTGAAGCAGTCAGGACCTGGCCTAGTGCAGCCCTCACAGAGCCTGTCCATCACCTGCACAGTCTCTGGTTTCTCATTAACTAACTATGGTGTACACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGAGTGGTGGAAACACAGACTATAATACACCTTTCACATCCAGACTGAGCATCAACAAGGACAATTCCAAGAGCCAAGTTTTCTTTAAAATGAACAGTCTGCAATCTAATGACACAGCCATATATTACTGTGCCAGAGCCCTCACCTACTATGATCTCGAGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTAGC

[0758] >Seq ID 269: Cetuximab N92F VL amino acid sequence

[0759] DILLTQSPVILSVSPGERVSFSC RASQSIGTNIH WYQQRTNGSPRLLIK YASESIS GIPSRFSGSGSGTDFTLSINSVESEDIADYYC QQNFNWPTT FGAGTKLELK

[0760] >Seq ID 270: Cetuximab N92F VL nucleotide sequence

[0761] GACATCTTGCTGACTCAGTCTCCAGTCATCCTGTCTGTGAGTCCAGGAGAAAGAGTCAGTTTCTCCTGCAGGGCCAGTCAGAGTATTGGCACAAACATACACTGGTATCAGCAAAGAACAAATGGTTCTCCAAGGCTTCTCATAAAGTATGCTTCTGAGTCTATCTCTGGGATTCCTTCCAGGTTTAGTGGCAGTGGATCAGGGACAGATTTTACTCTTAGCATCAACAGTGTGGAGTCTGAAGATATTGCAGATTATTACTGTCAACAAAATTTCAACTGGCCAACCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA

Claims

1. A bispecific tetravalent antibody having binding affinity for at least two epitopes of EGFR, comprising: An antibody framework, comprising: An antibody light chain having an antibody light chain variable (VL) domain, An antibody heavy chain having an antibody heavy chain variable (VH) domain, Wherein the antibody VL domain and the antibody VH domain form a Fab region, and A scFv domain having a scFv light chain variable (VL) domain and a scFv heavy chain variable (VH) domain, wherein the scFv domain is connected to at least one end of the antibody light chain or the antibody heavy chain through a domain linker.

2. The bispecific tetravalent antibody according to claim 1, wherein the two epitopes of EGFR comprise an EGFR wild-type (EGFRwt) epitope and an EGFRvIII epitope, and wherein the antibody has a stronger binding affinity for the EGFRvIII epitope than for the EGFRwt epitope.

3. The bispecific tetravalent antibody according to claim 2, wherein the antibody has a binding affinity with a first KD for the EGFRvIII epitope and a binding affinity with a second KD for the EGFRvIII epitope, wherein the first KD is higher than the second KD, and wherein the first KD is not less than 1E-11 M and the second KD is not greater than 1E-06 M.

4. The bispecific tetravalent antibody according to claim 2, wherein the EGFRwt epitope comprises an epitope having binding affinity for cetuximab, or wherein the EGFRvIII epitope comprises an epitope having binding affinity for ABT-806.

5. The bispecific tetravalent antibody according to claim 2, Wherein the scFv domain has binding affinity for the EGFRwt epitope and the Fab domain has binding affinity for the EGFRvIII epitope, or Wherein the scFv domain has binding affinity for the EGFRvIII epitope and the Fab domain has binding affinity for the EGFRwt epitope.

6. The bispecific tetravalent antibody according to claim 1, wherein the scFv domain is connected to the antibody light chain at its C-terminus, connected to the antibody light chain at its N-terminus, connected to the antibody heavy chain at its C-terminus, or connected to the antibody heavy chain at its N-terminus.

7. The bispecific tetravalent antibody according to claim 1, wherein the antibody framework comprises cetuximab, humanized cetuximab, de-immunized cetuximab or humanized de-immunized cetuximab, and wherein the scFv domain comprises a binding domain derived from the variable region of ABT-806 or humanized ABT-806.

8. The bispecific tetravalent antibody according to claim 7, wherein the antibody backbone comprises a de-matured mutation of cetuximab (numbered in sequence), and the de-matured mutation of cetuximab comprises VH-Y101A, VH-Y101W, VH-Y102A, VH-D103F, VH-D103W, VH-D103Y, VH-Y104L, VL-N92F, VL-N92K, or a combination thereof.

9. The bispecific tetravalent antibody according to claim 7, wherein the heavy chain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.1, 13, 79, 97, 101, 103, 105, 107, 123, 127, 129, 131, or 133, or wherein the light chain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.3, 81, 83, 99, 109, 125, or 135.

10. The bispecific tetravalent antibody according to claim 7, wherein the antibody backbone comprises humanized cetuximab, wherein the VH domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.237, or wherein the VL domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.

239.

11. The bispecific tetravalent antibody according to claim 7, wherein the antibody backbone comprises de-matured cetuximab, and wherein the VH domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.247, 251, 253, 259, 261, 263, 265, or 267, or wherein the VL domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.243 or 269.

12. The bispecific tetravalent antibody according to claim 7, wherein the antibody backbone comprises humanized de-matured cetuximab, and wherein the VH domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.245, 249, 255, or 257, or wherein the VL domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.

241.

13. The bispecific tetravalent antibody according to claim 7, wherein the scFv domain comprises the variable region of ABT-806 or humanized ABT-806.

14. The bispecific tetravalent antibody according to claim 7, wherein the scFv VH domain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.209, 225, 229, or 233, or wherein the scFv VL domain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.211, 227, 231, or 235.

15. The bispecific tetravalent antibody according to claim 7, wherein the VH domain of the antibody comprises 3 complementarity-determining regions (CDRs) of SEQ ID NO. 201, 237, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265 or 267; wherein the VL domain of the antibody comprises 3 CDRs of SEQ ID NO. 203, 239, 241, 243 or 269; wherein the scFv VH domain comprises 3 CDRs of SEQ ID NO. 209; and wherein the scFv VL domain comprises 3 CDRs of SEQ ID NO.

211.

16. The bispecific tetravalent antibody according to claim 1, wherein the antibody framework comprises ABT0806 or humanized ABT-806, and wherein the scFv domain comprises a binding domain of a variable region derived from cetuximab, humanized cetuximab, de-immunized cetuximab, humanized de-immunized cetuximab or nimotuzumab.

17. The bispecific tetravalent antibody according to claim 16, wherein the heavy chain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 9, 17, 19, 21, 23, 25, 27, 29, 31, 75, 93, 113, 115, 117, 119, 121, 137, 139, 141, 143, 145, 147 or 179, or wherein the light chain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 11, 33, 35, 37, 39, 41, 43, 77, 91, 95 or 111.

18. The bispecific tetravalent antibody according to claim 16, wherein the antibody framework comprises the ABT-806, and wherein the VH domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 209, or wherein the VL domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.

211.

19. The bispecific tetravalent antibody according to claim 16, wherein the antibody framework comprises the humanized ABT-806, and wherein the VH domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 225, 229 or 233, or wherein the antibody framework comprises the humanized ABT-806, and wherein the VL domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 227, 231 or 235.

20. The bispecific tetravalent antibody according to claim 16, wherein the scFv VH domain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 201, 205, 237, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265 or 267, or wherein the scFv VL domain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 203, 207, 239, 241, 243 or 269.

21. The bispecific tetravalent antibody according to claim 16, wherein the scFv domain comprises a binding domain derived from the variable region of the de - matured cetuximab or the humanized de - matured cetuximab, wherein the de - matured cetuximab or the humanized de - matured cetuximab comprises a cetuximab de - maturation mutation (numbered in sequence), and the cetuximab de - maturation mutation comprises VH - Y101A, VH - Y101W, VH - Y102A, VH - D103F, VH - D103W, VH - D103Y, VH - Y104L, VL - N92F, VL - N92K or a combination thereof.

22. The bispecific tetravalent antibody according to claim 16, wherein the antibody VH domain comprises 3 complementarity - determining regions (CDRs) of SEQ ID NO. 209; wherein the antibody VL domain comprises 3 CDRs of SEQ ID NO. 211; wherein the scFv VH domain comprises 3 CDRs of SEQ ID NO. 201, 205, 237, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265 or 267; and wherein the scFv VL domain comprises 3 CDRs of SEQ ID NO. 203, 207, 239, 241, 243 or 269.

23. The bispecific tetravalent antibody according to claim 1, wherein the antibody framework comprises nimotuzumab, and wherein the scFv domain comprises a binding domain derived from the variable region of ABT - 806 or humanized ABT - 806.

24. The bispecific tetravalent antibody according to claim 23, wherein the heavy chain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 5, 15, 45, 47, 49, 51, 53, 55 or 85, or wherein the light chain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 7, 57, 59, 61, 63, 65, 67, 69, 71 or 73.

25. The bispecific tetravalent antibody according to claim 23, wherein the VH domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 205, or wherein the VL domain of the antibody comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.

207.

26. The bispecific tetravalent antibody according to claim 23, wherein the scFv VH domain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 209, or wherein the scFv VL domain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO.

211.

27. A de - matured cetuximab monoclonal antibody, which comprises a light chain having a variable light (VL) domain and a heavy chain having a variable heavy (VH) domain, wherein the VL domain and the VH domain form a Fab region, and wherein the monoclonal antibody comprises a cetuximab de - maturation mutation selected from VH - Y101A, VH - Y101W, VH - Y102A, VH - D103F, VH - D103W, VH - D103Y, VH - Y104L, VL - N92F, VL - N92K or a combination thereof (numbered in sequence).

28. A humanized anti - EGFRvIII monoclonal antibody, which comprises a light chain having a variable light (VL) domain and a heavy chain having a variable heavy (VH) domain, wherein the VL domain and the VH domain form a Fab region, and wherein the heavy chain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 171, 149 or 177, or wherein the light chain comprises an amino acid sequence having at least 99% sequence identity with SEQ ID NO. 111, 173 or 175.

29. An isolated nucleic acid sequence encoding the bispecific tetravalent antibody according to claim 1.

30. A method for producing a bispecific tetravalent antibody, which comprises culturing a host cell to produce the bispecific antibody, wherein the host cell comprises the nucleic acid according to claim 1.

31. An immunoconjugate comprising the bispecific tetravalent antibody according to claim 1 and a cytotoxic agent, wherein the cytotoxic agent comprises a chemotherapeutic agent, a growth inhibitor, a toxin or a radioisotope.

32. A pharmaceutical composition comprising the bispecific tetravalent antibody according to claim 1 and a pharmaceutically acceptable carrier.

33. The pharmaceutical composition according to claim 32, which further comprises a radioisotope, a radionuclide, a toxin, a therapeutic agent, a chemotherapeutic agent or a combination thereof.

34. A pharmaceutical composition comprising the immunoconjugate according to claim 31 and a pharmaceutically acceptable carrier.

35. A method for treating a subject suffering from cancer, which comprises administering to the subject an effective amount of the bispecific tetravalent antibody according to claim 1, wherein the cancer comprises cells expressing EGFR, EGFRvIII or both.

36. The method according to claim 35, further comprising co-administering an effective amount of a therapeutic agent, wherein the therapeutic agent comprises an antibody, a chemotherapeutic agent, an enzyme, or a combination thereof.

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