Fc Variants With Optimized Fc Receptor Binding Properties

Abstract

The present invention relates to Fc variants with optimized Fc receptor binding properties, methods for their generation, Fc polypeptides comprising Fc variants with optimized Fc receptor binding properties, and methods for using Fc variants with optimized Fc receptor binding properties.

1 . An Fc variant of a parent Fc polypeptide comprising at least a first and a second substitution, said first and second substitutions each at a position selected from group consisting of 234, 235, 236, 239, 267, 268, 293, 295, 324, 327, 328, 330, and 332, wherein said Fc variant exhibits an increase in affinity for one or more receptors selected from the group consisting of FcγRI, FcγRIIa, and FcγRIIIa as compared to the increase in a affinity of said Fc variant for the FcγRIIb receptor, wherein the numbering is according to the EU index and wherein said increases in affinities are relative to said parent polypeptide.  
     
     
         2 . An Fc variant according to  claim 1 , wherein at least one of said substitutions is selected from the group consisting of 234G, 234I, 235D, 235E, 235I, 235Y, 236A, 236S, 239D, 267D, 267E, 267Q, 268D, 268E, 293R, 295E, 324G, 324I, 327H, 328A, 328F, 328I, 330I, 330L, 330Y, 332D, and 332E.  
     
     
         3 . An Fc variant according to  claim 2 , wherein said first and second substitutions are each selected from the group consisting of 234G, 234I, 235D, 235E, 235I, 235Y, 236A, 236S, 239D, 267D, 267E, 267Q, 268D, 268E, 293R, 295E, 324G, 324I, 327H, 328A, 328F, 328I, 330I, 330L, 330Y, 332D, and 332E.  
     
     
         4 . An Fc variant according to  claim 1 , wherein said Fc polypeptide further has increased affinity for FcγRI relative to the parent Fc polypeptide.  
     
     
         5 . An antibody or Fc fusion comprising an Fc variant according to  claim 1 .  
     
     
         6 . An Fc variant according to  claim 1 , wherein said modification is a reduced level of fucosylation relative to said parent Fc variant.  
     
     
         7 . An Fc variant according to  claim 1 , wherein said Fc variant mediates improved phagocytosis by FcγRIIa expressing cells relative to said parent Fc polypeptide.  
     
     
         8 . A composition comprising the Fc variant of  claim 1 , wherein said Fc variant comprises a glycosylated Fc region, wherein about 80-100% of the glycosylated Fc polypeptide in the composition comprises a mature core carbohydrate structure with no fucose.  
     
     
         9 . An Fc variant of a parent Fc polypeptide comprising at least a first and a second substitution, said first and second substitutions each at a position selected from group consisting of 234, 235, 236, 239, 267, 268, 293, 295, 324, 327, 328, 330, and 332, wherein said Fc variant exhibits an increase in a affinity of said Fc variant for the FcγRIIb receptor as compared to the increase in affinity for one or more receptors selected from the group consisting of FcγRI, FcγRIIa, and FcγRIIIa, wherein the numbering is according to the EU index and wherein said increases in affinities are relative to said parent polypeptide.  
     
     
         10 . An Fc variant according to  claim 9 , wherein at least one of said first and second substitutions is selected from the group consisting of 236A, 236S, 239D, 267D, 267E, 267Q, 268D, 268E, 293R, 295E, 324G, 324I, 327H, 328A, 328F, 330I, 330L, 330Y, 332D, and 332E.  
     
     
         11 . An Fc variant according to  claim 10 , wherein each of said first and second substitutions is selected from the group consisting of 236A, 236S, 239D, 267D, 267E, 267Q, 268D, 268E, 293R, 295E, 324G, 324I, 327H, 328A, 328F, 330I, 330L, 330Y, 332D, and 332E.  
     
     
         12 . An Fc variant comprising a first substitution at a position selected from the group consisting of 234, 235, 236, 239, 267, 268, 293, 295, 324, 327, 328, 330, and 332, and a second substitution selected from the group consisting of 247L, 255L, 270E, 280H, 280Q, 280Y, 298A, 298T, 392T, 396L, 326A, 326D, 326E, 326W, 333A, 334A, 334L, and 421 K.  
     
     
         13 . The Fc variant according to  claim 12 , said substitution comprising at least two amino acids positions selected from the group consisting of 235, 236, 237, 238, 239, 265, 266, 267, 269, 270, 295, 296, 298, 299, 325, 326, 327, 328, 329, 330, and 332.  
     
     
         14 . An Fc variant comprising a first substitution at a position selected from the group consisting of 239 and 332, and a second substitution at a position selected from the group consisting of 233, 234, 241, 264, 265, 268, 328, 333 and 334.  
     
     
         15 . An Fc variant according to  claim 14  further comprising a substitution at position 239 and position 332.  
     
     
         16 . An Fc variant according to  claim 14 , wherein said first substitution is selected from the group consisting of 239D and 332E.  
     
     
         17 . An Fc variant according to  claim 14 , wherein said second substitution is selected from the group consisting of 233H, 234K, 241H, 241Q, 241R, 264T, 265N, 265K, 265H, 265Q, 265G, 265S, 265L, 268E, 328K, 333T, 333H, and 334R.  
     
     
         18 . A method of activating an receptor selected from the group consisting of FcγRI, FcγRIIa, and FcγRIIIa relative to FcγRIIb receptor, said method comprising contacting a cell comprising a receptor selected from the group consisting of FcγRI, FcγRIIa, and FcγRIIIa with an Fc variant according to  claim 1 .  
     
     
         19 . A method of activating an FcγRIIb receptor relative to a receptor selected from the group consisting of FcγRI, FcγRIIa, and FcγRIIIa, said method comprising contacting a cell comprising a receptor selected from the group consisting of FcγRI, FcγRIIa, and FcγRIIIa with an Fc variant according to  claim 9 .  
     
     
         20 . An Fc variant of a parent mouse Fc polypeptide, said Fc variant comprising a substitution at a position selected from the group consisting of 236, 239, 268, 330, and 332.  
     
     
         21 . An Fc variant according to  claim 19 , wherein said substitution is selected from the group consisting of 236A, 239D, 268E, 330Y, and 332E.