Human antibodies against tissue factor and methods of use thereof

Abstract

Isolated human monoclonal antibodies which bind to human TF and related antibody-based compositions and molecules, are disclosed. Also disclosed are pharmaceutical compositions comprising the antibodies, and therapeutic and diagnostic methods for using the antibodies.

1 . A human antibody which binds human Tissue Factor. 
     
     
         2 - 9 . (canceled) 
     
     
         10 . The antibody of  claim 1 , wherein said antibody competes for Tissue Factor binding with an antibody comprising a VH region comprising the sequence of SEQ ID NO:9 and a VL region comprising the sequence of SEQ ID NO:65. 
     
     
         11 . (canceled) 
     
     
         12 . The antibody of  claim 1 , wherein said antibody competes for Tissue Factor binding with an antibody comprising a VH region comprising the sequence of SEQ ID NO:37 and a VL region comprising the sequence of SEQ ID NO:93. 
     
     
         13 - 27 . (canceled) 
     
     
         28 . The antibody of  claim 1 , wherein the antibody comprises:
 a) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:10, 11 and 12 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:66, 67 and 68,   b) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:14, 15 and 16 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:70, 71 and 72,   c) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:18, 19, 20 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:74, 75 and 76,   d) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:22, 23 and 24 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:78, 79 and 80,   e) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO: 26, 27 and 28 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO: 82, 83 and 84, or   f) a variant of any of said antibodies, wherein said variant preferably has at most 1, 2 or 3 amino-acid modifications, more preferably amino-acid substitutions, such as conservative amino-acid substitutions in said sequences.   
     
     
         29 - 33 . (canceled) 
     
     
         34 . The antibody of  claim 1 , wherein the antibody comprises:
 a) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:6, 7 and 8 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:62, 63 and 64,   b) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:50, 51 and 52 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:106, 107 and 108, or   c) a variant of any of said antibodies, wherein said variant preferably has at most 1, 2 or 3 amino-acid modifications, more preferably amino-acid substitutions, such as conservative amino-acid substitutions in said sequences.   
     
     
         35 - 39 . (canceled) 
     
     
         40 . The antibody of  claim 1 , wherein the antibody comprises:
 a) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO: 30, 31 and 32 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO: 86, 87 and 88,   b) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:34, 35 and 36 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:90, 91 and 92,   c) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:38, 39 and 40 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:94, 95 and 96,   d) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:54, 55 and 56 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:110, 11 and 112, or   e) a variant of any of said antibodies, wherein said variant preferably has at most 1, 2 or 3 amino-acid modifications, more preferably amino-acid substitutions, such as conservative amino-acid substitutions in said sequences.   
     
     
         41 - 45 . (canceled) 
     
     
         46 . The antibody of  claim 1 , wherein the antibody comprises:
 a) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:2, 3 and 4 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:58, 59 and 60,   b) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:42, 43 and 44 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:98, 99 and 100,   c) a VH region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:46, 47 and 48 and a VL region comprising the CDR1, 2 and 3 sequences of SEQ ID NO:102, 103 and 104, or   d) a variant of any of said antibodies, wherein said variant preferably has at most 1, 2 or 3 amino-acid modifications, more preferably amino-acid substitutions, such as conservative amino-acid substitutions in said sequences.   
     
     
         47 - 55 . (canceled) 
     
     
         56 . The antibody of  claim 1  which competes for Tissue Factor binding with an antibody comprising
 a) a VH region comprising the sequence of SEQ ID NO:9 and a VL region comprising the sequence of SEQ ID NO: 65, 
 b) a VH region comprising the sequence of SEQ ID NO:1 and a VL region comprising the sequence of SEQ ID NO:57, 
 c) a VH region comprising the sequence of SEQ ID NO:5 and a VL region comprising the sequence of SEQ ID NO:61, 
 d) a VH region comprising the sequence of SEQ ID NO:13 and a VL region comprising the sequence of SEQ ID NO:69, 
 e) a VH region comprising the sequence of SEQ ID NO:17 and a VL region comprising the sequence of SEQ ID NO:73, 
 f) a VH region comprising the sequence of SEQ ID NO:21 and a VL region comprising the sequence of SEQ ID NO:77, 
 g) a VH region comprising the sequence of SEQ ID NO:25 and a VL region comprising the sequence of SEQ ID NO:81, 
 h) a VH region comprising the sequence of SEQ ID NO:29 and a VL region comprising the sequence of SEQ ID NO:85, 
 i) a VH region comprising the sequence of SEQ ID NO:33 and a VL region comprising the sequence of SEQ ID NO:89, 
 j) a VH region comprising the sequence of SEQ ID NO:37 and a VL region comprising the sequence of SEQ ID NO:93, 
 k) a VH region comprising the sequence of SEQ ID NO:41 and a VL region comprising the sequence of SEQ ID NO: 97, 
 l) a VH region comprising the sequence of SEQ ID NO:45 and a VL region comprising the sequence of SEQ ID NO:101, 
 m) a VH region comprising the sequence of SEQ ID NO:49 and a VL region comprising the sequence of SEQ ID NO:105, or 
 n) a VH region comprising the sequence of SEQ ID NO:53 and a VL region comprising the sequence of SEQ ID NO:109. 
 
     
     
         57 . (canceled) 
     
     
         58 . The antibody of  claim 1 , wherein the antibody comprises:
 a heavy chain variable region derived from a human germline V H  sequence selected from the group consisting of: IGHV1-18*01, IGHV3-23*01, IGHV3-30*01, IGHV3-33*01, IGHV3-33*03, IGHV1-69*02, IGHV1-69*04 and IGHV5-51*01   
       and/or
 a light chain variable region derived from a human germline Vκ sequence selected from the group consisting of: IGKV3-20*01, IGKV1-13*02, IGKV3-11*01, and IGKV1D-16*01. 
 
     
     
         59 . (canceled) 
     
     
         60 . The antibody of  claim 28 , wherein the antibody is conjugated to another moiety, such as a cytotoxic moiety, a radioisotope or a drug. 
     
     
         61 - 63 . (canceled) 
     
     
         64 . The antibody of  claim 28 , wherein said antibody is a monovalent antibody, wherein said monovalent antibody is constructed by a method comprising:
 i) providing a nucleic acid construct encoding the light chain of said monovalent antibody, said construct comprising a nucleotide sequence encoding the VL region of a selected antigen specific antibody and a nucleotide sequence encoding the constant CL region of an Ig, wherein said nucleotide sequence encoding the VL region of a selected antigen specific antibody and said nucleotide sequence encoding the CL region of an Ig are operably linked together, and wherein, in case of an IgG1 subtype, the nucleotide sequence encoding the CL region has been modified such that the CL region does not contain any amino acids capable of forming disulfide bonds or covalent bonds with other peptides comprising an identical amino acid sequence of the CL region in the presence of polyclonal human IgG or when administered to an animal or human being;   ii) providing a nucleic acid construct encoding the heavy chain of said monovalent antibody, said construct comprising a nucleotide sequence encoding the VH region of a selected antigen specific antibody and a nucleotide sequence encoding a constant CH region of a human Ig, wherein the nucleotide sequence encoding the CH region has been modified such that the region corresponding to the hinge region and, as required by the Ig subtype, other regions of the CH region, such as the CH3 region, does not comprise any amino acid residues which participate in the formation of disulphide bonds or covalent or stable non-covalent inter-heavy chain bonds with other peptides comprising an identical amino acid sequence of the CH region of the human Ig in the presence of polyclonal human IgG or when administered to an animal human being, wherein said nucleotide sequence encoding the VH region of a selected antigen specific antibody and said nucleotide sequence encoding the CH region of said Ig are operably linked together;   iii) providing a cell expression system for producing said monovalent antibody;   iv) producing said monovalent antibody by co-expressing the nucleic acid constructs of (i) and (ii) in cells of the cell expression system of (iii).   
     
     
         65 - 66 . (canceled) 
     
     
         67 . A bispecific molecule comprising an antibody of  claim 28  and a second binding specificity, such as a binding specificity for a human effector cell, a human Fc receptor or a T cell receptor. 
     
     
         68 . An expression vector comprising a nucleotide sequence encoding one or more of the amino acid sequences selected from the group consisting of SEQ ID NO:1-SEQ ID NO: 112. 
     
     
         69 . (canceled) 
     
     
         70 . A recombinant eukaryotic or prokaryotic host cell which produces an antibody as defined in  claim 28 . 
     
     
         71 . A pharmaceutical composition comprising an antibody as defined in  claim 28 , and a pharmaceutically acceptable carrier. 
     
     
         72 - 83 . (canceled) 
     
     
         84 . A method for inhibiting growth and/or proliferation of a tumor cell expressing Tissue Factor, comprising administration, to an individual in need thereof, of an antibody of  claim 28 . 
     
     
         85 . A method for producing an antibody, said method comprising the steps of
 a) culturing a host cell of  claim 70 , and   b) purifying the antibody from the culture media.   
     
     
         86 . A diagnostic composition comprising an antibody as defined in  claim 28 . 
     
     
         87 . A method for detecting the presence of Tissue Factor in a sample, comprising:
 contacting the sample with an antibody of  claim 28  under conditions that allow for formation of a complex between the antibody or bispecific molecules and Tissue Factor; and   analyzing whether a complex has been formed.   
     
     
         88 . A kit for detecting the presence of Tissue Factor in a sample comprising
 an antibody of  claim 28 ; and   instructions for use of the kit.