Optimized Fc Variants and Methods for Their Generation

Abstract

The present invention relates to optimized Fc variants, methods for their generation, and antibodies and Fc fusions comprising optimized Fc variants.

1 . An antibody or immunoadhesin of a parent Fc polypeptide, said antibody or immunoadhesin comprising an amino acid substitution at position 267, wherein said antibody or immunoadhesin alters binding affinity to an FcγR as compared to said parent Fc polypeptide, and wherein numbering is according to the EU index. 
     
     
         2 . An antibody or immunoadhesin of a parent Fc polypeptide, said antibody or immunoadhesin comprising an amino acid substitution selected from the group consisting of 267T, 267H, 267D, 267N, 267E, 267Q, 267K, 267R, 267V, 267L, 267I, 267F, 267M, 267Y, 267W and 267P, wherein numbering is according to the EU index. 
     
     
         3 . An antibody or immunoadhesin of a parent Fc polypeptide, said antibody or immunoadhesin comprising an amino acid substitution at position 267, wherein said antibody or immunoadhesin further comprises an amino acid substitution at a position selected from the group consisting of 221, 222, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 258, 262, 263, 264, 265, 266, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 280, 281, 283, 285, 286, 288, 290, 291, 293, 294, 295, 296, 297, 298, 299, 300, 302, 313, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335 336 and 428, wherein said antibody or immunoadhesin increases binding affinity to an FcγR as compared to said parent Fc polypeptide, and wherein numbering is according to the EU index. 
     
     
         4 . An antibody or immunoadhesin according to  claim 3  wherein said antibody or immunoadhesin comprises an amino acid substitution at position 267T, 267H, 267D, 267N, 267E, 267Q, 267K, 267R, 267V, 267L, 267I, 267F, 267M, 267Y, 267W and 267P, wherein said antibody or immunoadhesin further comprises an amino acid substitution at a position selected from the group consisting of 221, 222, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 258, 262, 263, 264, 265, 266, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 280, 281, 283, 285, 286, 288, 290, 291, 293, 294, 295, 296, 297, 298, 299, 300, 302, 313, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335 336 and 428. 
     
     
         5 . An antibody or immunoadhesin according to  claim 4  wherein said antibody or immunoadhesin is selected from the group consisting of 267Q/327S and 267L/327A. 
     
     
         6 . An antibody or immunoadhesin according to  claim 2  wherein said antibody or immunoadhesin increases binding affinity to an FcγR as compared to said parent polypeptide, wherein numbering is according to the EU index. 
     
     
         7 . An antibody or immunoadhesin according to  claim 6  wherein said FcγR is FcγRIIIa. 
     
     
         8 . An antibody or immunoadhesin according to  claim 7  wherein said FcγRIlIa is a V158 or F158 allotype of FcγRIIIa. 
     
     
         9 . An antibody or immunoadhesin according to  claim 2  wherein said antibody or immunoadhesin decreases binding affinity to an FcγR as compared to said parent polypeptide, wherein numbering is according to the EU index. 
     
     
         10 . An antibody or immunoadhesin according to  claim 1  wherein said antibody or immunoadhesin is an antibody. 
     
     
         11 . An antibody according to  claim 10  wherein said antibody is selected from the group consisting of a human antibody, a humanized antibody, a monoclonal antibody and an antibody fragment. 
     
     
         12 . An antibody or immunoadhesin according to  claim 1  wherein said antibody or immunoadhesin further comprises an engineered glycoform. 
     
     
         13 . An antibody or immunoadhesin according to  claim 1  wherein said antibody or immunoadhesin has specificity for a target antigen selected from the group consisting of CD19, CD20, CD22, CD30, CD33, CD40, CD40L, CD52, Her2/neu, EGFR, EpCAM, MUC1, GD3, CEA, CA 125, HLA-DR, TNFalpha, MUC18, prostate specific membrane antigen (PMSA) and VEGF. 
     
     
         14 . A composition comprising the antibody or immunoadhesin according to  claim 1  further comprising a pharmaceutically acceptable carrier. 
     
     
         15 . A method of treating a mammal in need of said treatment, comprising administering an antibody or immunoadhesin of a parent Fc polypeptide, said antibody or immunoadhesin comprising an amino acid substitution at position 267, wherein said antibody or immunoadhesin alters binding affinity to an FcγR as compared to said parent Fc polypeptide, and wherein numbering is according to the EU index. 
     
     
         16 . A method according to  claim 15  wherein said antibody or immunoadhesin comprises an amino acid substitution selected from the group consisting of 267T, 267H, 267D, 267N, 267E, 267Q, 267K, 267R, 267V, 267L, 267I, 267F, 267M, 267Y, 267W and 267P. 
     
     
         17 . A method according to  claim 15  wherein said antibody or immunoadhesin further comprises an amino acid substitution at a position selected from the group consisting of 221, 222, 224, 227, 228, 230, 231, 223, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 258, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 280, 281, 283, 285, 286, 288, 290, 291, 293, 294, 295, 296, 297, 298, 299, 300, 302, 313, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335 336 and 428. 
     
     
         18 . A method according to  claim 15  wherein said antibody or immunoadhesin is selected from the group consisting of 267E/328F, 267Q/327S and 267L/327A. 
     
     
         19 . A method according to  claim 15  wherein said FcγR is FcγRIIIa. 
     
     
         20 . A method according to  claim 19  wherein said FcγRIlIla is a V158 or F158 allotype of FcγRIIIa. 
     
     
         21 . A method according to  claim 15  wherein said antibody or immunoadhesin is an antibody. 
     
     
         22 . A method according to  claim 21  wherein said antibody is selected from the group consisting of a human antibody, a humanized antibody, a monoclonal antibody and an antibody fragment. 
     
     
         23 . A method according to  claim 15  wherein said antibody or immunoadhesin further comprises an engineered glycoform. 
     
     
         24 . A method according to  claim 15  wherein said antibody or immunoadhesin has specificity for a target antigen selected from the group consisting of CD19, CD20, CD22, CD30, CD33, CD40, CD40L, CD52, Her2/neu, EGFR, EpCAM, MUC1, GD3, CEA, CA 125, HLA-DR, TNFalpha, MUC18, prostate specific membrane antigen (PMSA) and VEGF.