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 . 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 chain frameworks, which humanized immunoglobulin specifically binds to an antigen with a binding affinity of at least 10 8  M −1 , 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:  
     
     
         25 . 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.  
     
     
         26 . A pharmaceutical composition comprising a humanized immunoglobulin of  claim 24  in a pharmaceutically acceptable carrier.  
     
     
         27 . A method of producing a humanized immunoglobulin, comprising the steps of: 
 (1) comparing the sequence of a donor immunoglobulin heavy chain variable region against a collection of sequences of human heavy chain variable regions;    (2) selecting a human heavy chain variable region from the collection of human heavy chain variable regions to provide an acceptor heavy chain variable region, and to maximise homology with said donor sequence;    (3) synthesizing a DNA segment encoding a humanized heavy chain variable region, comprising CDRs from the donor immunoglobulin heavy chain variable region and a variable region framework from the selected acceptor heavy chain variable region;    (4) introducing the DNA segment encoding the humanized immunoglobulin heavy chain variable region and a DNA segment encoding a humanized immunoglobulin light chain variable region into a cell; and    (5) expressing the DNA segments in the cell to produce the humanized immunoglobulin.    
     
     
         28 . A method of producing a humanized immunoglobulin, comprising the steps of: 
 (1) comparing the sequence of a donor immunoglobulin light chain variable region against a collection of sequences of human light chain variable regions;    (2) selecting a human light chain variable region from the collection of human light chain variable regions to provide an acceptor light chain variable region, and to maximise homology with said donor sequence;    (3) synthesizing a DNA segment encoding a humanized light chain variable region, comprising CDRs from the donor immunoglobulin light chain variable region and a variable region framework from the selected acceptor light chain variable region;    (4) introducing the DNA segment encoding the humanized immunoglobulin light chain variable region and a DNA segment encoding a humanized immunoglobulin heavy chain variable region into a cell; and    (5) expressing the DNA segments in the cell to produce the humanized immunoglobulin.    
     
     
         29 . 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 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.  
     
     
         30 . 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 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 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.