Antibodies binding axl

Abstract

The present invention relates to anti-AXL antibodies, immunoconjugates, compositions and method of treatment of cancer with such anti-AXL antibodies, immunoconjugates, or compositions.

1 - 62 . (canceled) 
     
     
         63 . A nucleic acid encoding an antibody which binds to AXL, wherein the antibody does not compete for AXL binding with Growth Arrest-Specific 6 (Gas6). 
     
     
         64 . The nucleic acid according to  claim 63 , wherein maximal binding of the antibody to AXL in the presence of Gas6 is at least 90% of binding in the absence of Gas6 as determined by the method disclosed in Example 2, or as determined by a competition assay, wherein competition between the antibody binding to AXL and Gas6 is determined on A431 cells preincubated with Gas6 and without Gas6. 
     
     
         65 . The nucleic acid according to  claim 63 , wherein (a) the antibody has a binding affinity (K D ) in the range of 0.3×10 −9  to 63×10 −9  M to human AXL, and wherein said binding affinity is measured using a Bio-layer Interferometry using soluble AXL extracellular domain, and/or (b) wherein the antibody has a dissociation rate of 9.7×10 −5  to 4.4×10 −3  s −1  to human AXL, and wherein the dissociation rate is measured by Bio-layer Interferometry using soluble recombinant AXL extracellular domain. 
     
     
         66 . The nucleic acid according to  claim 63 , wherein the antibody comprises at least one binding region comprising a pair of variable heavy chain (VH) and variable light chain (VL) regions selected from the group consisting of:
 a) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 36, 37, and 38, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 39, GAS, and 40, respectively;   b) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 46, 47, and 48, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 49, AAS, and 50, respectively;   c) a VH region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 114, 115, and 116, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences of SEQ ID NOs: 117, DAS, and 118, respectively;   d) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 51, 52, and 53, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 55, GAS, and 56, respectively;   e) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 51, 52, and 54, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 55, GAS, and 56, respectively;   f) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 57, 58, and 59, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 60, GAS, and 61, respectively;   g) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 62, 63, and 64, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 65, GAS, and 66, respectively;   h) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 67, 68, and 69, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 70, GAS, and 71, respectively;   i) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 72, 73, and 75, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 76, ATS, and 77, respectively;   j) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 72, 74, and 75, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 76, ATS, and 77, respectively;   k) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 78, 79, and 80, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 81, AAS, and 82, respectively;   l) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 83, 84, and 85, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 86, GAS, and 87, respectively;   m) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 88, 89, and 90, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 91, GAS, and 92, respectively;   n) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 93, 94, and 95, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 96, GAS, and 97, respectively;   o) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 98, 99, and 100, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 101, DAS, and 102, respectively;   p) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 103, 104, and 105, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 106, GAS, and 107, respectively;   q) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 108, 109, and 110, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 112, AAS, and 113, respectively;   r) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 108, 109, and 111, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 112, AAS, and 113, respectively;   s) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 41, 42, and 43, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 44, AAS, and 45, respectively;   t) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 93, 94, and 95, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 128, XAS, wherein X is D or G, and 129, respectively;   u) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 46, 119, and 120, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 49, AAS, and 50, respectively;   v) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 123, 124, and 125, respectively, and a VL region comprising CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 60, GAS, and 61, respectively;   w) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 121, 109, and 122, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 112, AAS, and 113, respectively; and   x) a VH region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs:93, 126, and 127, respectively, and a VL region comprising the CDR1, CDR2, and CDR3 sequences set forth in SEQ ID NOs: 96, GAS, and 97.   
     
     
         67 . The nucleic acid according to  claim 63 , wherein the antibody comprises at least one binding region comprising a pair of variable heavy chain (VH) and variable light chain (VL) regions selected from the group consisting of:
 a) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 1 and 2, respectively;   b) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 5 and 6, respectively;   c) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 34 and 35, respectively;   d) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 7 and 9, respectively;   e) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 10 and 11, respectively;   f) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 16 and 18, respectively;   g) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 25 and 26, respectively;   h) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 31 and 33, respectively;   i) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 3 and 4, respectively;   j) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 8 and 9, respectively;   k) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 12 and 13, respectively;   l) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 14 and 15, respectively;   m) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 17 and 18, respectively;   n) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 19 and 20, respectively;   o) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 21 and 22, respectively;   p) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 23 and 24, respectively;   q) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 27 and 28, respectively;   r) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 29 and 30, respectively; and   s) VH and VL regions comprising the amino acid sequences of SEQ ID NOs: 32 and 33, respectively.   
     
     
         68 . The nucleic acid according to  claim 63 , wherein the antibody comprises a heavy chain of an isotype selected from the group consisting of IgG1, IgG2, IgG3, and IgG4. 
     
     
         69 . The nucleic acid according to  claim 63 , wherein the antibody is a full length monoclonal antibody. 
     
     
         70 . The nucleic acid according to  claim 63 , wherein the antibody is a single-chain antibody. 
     
     
         71 . The nucleic acid according to  claim 63 , wherein the antibody is an effector-function deficient antibody, a stabilized IgG4 antibody, or a monovalent antibody. 
     
     
         72 . The nucleic acid according to  claim 63 , wherein the antibody is a bispecific antibody. 
     
     
         73 . A nucleic acid construct encoding at least one amino acid sequence selected from the group consisting of SEQ ID Nos.: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, and 135. 
     
     
         74 . A nucleic acid construct encoding a pair of amino acid sequences selected from the group consisting of:
 a) SEQ ID Nos.: 1 and 2,   b) SEQ ID Nos.: 3 and 4,   c) SEQ ID Nos.: 5 and 6,   d) SEQ ID Nos.: 7 and 9,   e) SEQ ID Nos.: 8 and 9,   f) SEQ ID Nos.: 10 and 11,   g) SEQ ID Nos.: 12 and 13,   h) SEQ ID Nos.: 14 and 15,   i) SEQ ID Nos.: 16 and 18,   j) SEQ ID Nos.: 17 and 18,   k) SEQ ID Nos.: 19 and 20,   l) SEQ ID Nos.: 21 and 22,   m) SEQ ID Nos.: 23 and 24,   n) SEQ ID Nos.: 25 and 26,   o) SEQ ID Nos.: 27 and 28,   p) SEQ ID Nos.: 29 and 30,   q) SEQ ID Nos.: 31 and 33,   r) SEQ ID Nos.: 31 and 32,   s) SEQ ID Nos.: 33 and 35, and   t) SEQ ID Nos.: 136 and 140.   
     
     
         75 . A nucleic acid construct encoding a heavy chain variable region or light chain variable region comprising three CDR sequences selected from the group consisting of:
 a) SEQ ID Nos.: 36, 37 and 38,   b) SEQ ID Nos.: 41, 42 and 43,   c) SEQ ID Nos.: 46, 47 and 48,   d) SEQ ID Nos.: 51, 52 and 53,   e) SEQ ID Nos.: 51, 52 and 54,   f) SEQ ID Nos.: 57, 58 and 59,   g) SEQ ID Nos.: 62, 63 and 64,   h) SEQ ID Nos.: 67, 68 and 69,   i) SEQ ID Nos.: 72, 73 and 74,   j) SEQ ID Nos.: 72, 73 and 75,   k) SEQ ID Nos.: 78, 79 and 80,   l) SEQ ID Nos.: 83, 84 and 85,   m) SEQ ID Nos.: 88, 89 and 90,   n) SEQ ID Nos.: 93, 94 and 95,   o) SEQ ID Nos.: 98, 99 and 100,   p) SEQ ID Nos.: 103, 104 and 105,   q) SEQ ID Nos.: 108, 109 and 110,   r) SEQ ID Nos.: 108, 109 and 111,   s) SEQ ID Nos.: 114, 115 and 116,   t) SEQ ID Nos.: 137, 138 and 139,   u) SEQ ID No.: 39, GAS and SEQ ID No.: 32,   v) SEQ ID No.: 44, AAS and SEQ ID No.: 45,   w) SEQ ID No.: 49, AAS and SEQ ID No.: 50,   x) SEQ ID No.: 55, GAS and SEQ ID No.: 56,   y) SEQ ID No.: 60, GAS and SEQ ID No.: 61,   z) SEQ ID No.: 65, GAS and SEQ ID No.: 66,   aa) SEQ ID No.: 70, GAS and SEQ ID No.: 71,   bb) SEQ ID No.: 76, ATS and SEQ ID No.: 77,   cc) SEQ ID No.: 81, AAS and SEQ ID No.: 82,   dd) SEQ ID No.: 86, GAS and SEQ ID No.: 87,   ee) SEQ ID No.: 91, GAS and SEQ ID No.: 92,   ff) SEQ ID No.: 96, GAS and SEQ ID No.: 97,   gg) SEQ ID No.: 101, DAS and SEQ ID No.: 102,   hh) SEQ ID No.: 106, GAS and SEQ ID No.: 107,   ii) SEQ ID No.: 117, DAS and SEQ ID No.: 118, and   jj) SEQ ID No.: 141, AAS and SEQ ID No.: 142.   
     
     
         76 . An expression vector comprising the nucleic acid according to  claim 63 . 
     
     
         77 . An expression vector comprising the nucleic acid construct according to  claim 73 . 
     
     
         78 . A host cell comprising the vector according to  claim 76 . 
     
     
         79 . A host cell comprising the vector according to  claim 77 . 
     
     
         80 . A method for producing an antibody which binds to AXL, wherein the antibody does not compete for AXL binding with Growth Arrest-Specific 6 (Gas6), said method comprising the steps of:
 a) culturing a host cell according to  claim 78 , and   b) purifying the antibody from the culture media.   
     
     
         81 . A method for producing an antibody which binds to AXL, wherein the antibody does not compete for AXL binding with Growth Arrest-Specific 6 (Gas6), said method comprising the steps of:
 a) culturing a host cell according to  claim 79 , and   b) purifying the antibody from the culture media.   
     
     
         82 . A method of detecting the presence of AXL antibody, or a cell expressing AXL, in a sample comprising the steps of;
 a) contacting the sample with an antibody which binds to AXL, wherein the antibody does not compete for AXL binding with the ligand Growth Arrest-Specific 6 (Gas6), under conditions that allow for formation of a complex between the antibody and AXL; and   b) analyzing whether a complex has been formed.