Humanised antibodies

Abstract

CDR-grafted antibody heavy and light chains comprise acceptor framework and donor antigen binding regions, the heavy chains comprising donor residues at at least one of positions (6, 23) and/or (24, 48) and/or (49, 71) and/or (73, 75) and/or (76) and/or (78) and (88) and/or (91). The CDR-grafted light chains comprise donor residues at at least one of positions (1) and/or (3) and (46) and/or (47) or at at least one of positions (46, 48, 58) and (71). The CDR-grafted antibodies are preferably humanised antibodies, having non human, e.g. rodent, donor and human acceptor frameworks, and may be used for in vivo therapy and diagnosis. A generally applicable protocol is disclosed for obtaining CDR-grafted antibodies.

1 - 23 . (canceled)  
     
     
         24 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) from a donor immunoglobulin and heavy and light chain variable region frameworks from acceptor immunoglobulin heavy and light chain frameworks, which humanized immunoglobulin specifically binds to an antigen with an affinity constant of at least about 10 8  M −1  and a binding affinity similar to that of the donor immunoglobulin, wherein the sequence of the acceptor immunoglobulin heavy chain variable region framework is a consensus sequence of human immunoglobulin heavy chain variable region frameworks.  
     
     
         25 . A vector comprising first and second polynucleotides according to  claim 24 .  
     
     
         26 . A cell line transfected with a vector according to  claim 25 .  
     
     
         27 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) from a donor immunoglobulin and heavy and light chain variable region frameworks from human acceptor immunoglobulin heavy and light chains, which humanized immunoglobulin specifically binds to an antigen with an affinity constant of at least about 10 8  M −1  and a binding affinity similar to that of the donor immunoglobulin, wherein said humanized immunoglobulin heavy chain comprises one or more amino acids from the donor immunoglobulin heavy chain framework outside the Kabat CDRs and the structural loop CDRs of the variable regions, wherein the donor amino acids substitute for corresponding amino acids in the acceptor immunoglobulin heavy chain framework, and each of these said donor amino acids is adjacent to a CDR in the donor immunoglobulin sequence or contributes to antigen binding as determined by X-ray crystallography.  
     
     
         28 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) from a donor immunoglobulin and heavy and light chain variable region frameworks from acceptor immunoglobulin heavy and light chain frameworks, which humanized immunoglobulin specifically binds to an antigen with a binding affinity similar to that of the donor immunoglobulin, wherein the sequence of the acceptor immunoglobulin heavy chain variable region framework is a consensus sequence of human immunoglobulin heavy chain variable region frameworks.  
     
     
         29 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) from a donor immunoglobulin and heavy and light chain variable region frameworks from human acceptor immunoglobulin heavy and light chains, which humanized immunoglobulin specifically binds to an antigen with an affinity constant of at least about 10 8  M −1  and a binding affinity similar to that of the donor immunoglobulin, wherein the sequence of the humanized immunoglobulin heavy chain variable region framework has 66 variable region framework residues identical to the variable region framework residues of the donor immunoglobulin heavy chain variable region framework and at least 74 residues identical to an acceptor human immunoglobulin heavy chain variable region amino acid sequence.  
     
     
         30 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) from a donor immunoglobulin and heavy and light chain variable region frameworks from human acceptor immunoglobulin heavy and light chains, which humanized immunoglobulin specifically binds to an antigen with a binding affinity similar to that of the donor immunoglobulin, wherein the sequence of the humanized immunoglobulin heavy chain variable region framework has 66 variable region framework residues identical to the variable region framework residues of the donor immunoglobulin heavy chain variable region framework and at least 74 residues identical to an acceptor human immunoglobulin heavy chain variable region amino acid sequence.  
     
     
         31 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) from a donor immunoglobulin and heavy and light chain variable region frameworks from human acceptor immunoglobulin heavy and light chains, which humanized immunoglobulin specifically binds to an antigen with a binding affinity similar to that of the donor immunoglobulin, wherein said humanized immunoglobulin heavy chain comprises one or more amino acids from the donor immunoglobulin heavy chain framework outside the Kabat CDRs and the structural loop CDRs of the variable regions, and each of said donor amino acids is adjacent to a CDR in the donor immunoglobulin sequence or contributes to antigen binding as determined by X-ray crystallography.  
     
     
         32 . First and second polynucleotides respectively encoding heavy and light chain variable regions of a humanized immunoglobulin having complementarity determining regions (CDRs) in and heavy and light chain variable region frameworks from human acceptor immunoglobulin heavy and light chains, which humanized immunoglobulin specifically binds to an antigen with a binding affinity similar to that of the donor immunoglobulin, wherein said humanized immunoglobulin heavy chain comprises one or more amino acids from the donor immunoglobulin heavy chain framework outside the Kabat CDRs and the structural loops of the variable region, and wherein the donor amino acids substitute for corresponding amino acids in the acceptor immunoglobulin heavy chain framework.