Host cell for making antibody fc-heterodimeric molecules using electrostatic steering effects

Abstract

The invention relates to methods of making Fc-heterodimeric proteins or polypeptides. The invention also relates to the Fc-heterodimeric proteins or polypeptides themselves, including the individual polypeptide components that comprise the heterodimer Nucleic acids encoding such polypeptides, expression vectors, and host cells. Moreover, the invention relates to pharmaceutical compositions comprising one of more Fc-heterodimeric proteins or polypeptides.

1 . A host cell for producing a heterodimeric protein, said host cell comprising a nucleic acid encoding a first CH3-containing polypeptide and a nucleic acid encoding a second CH3-containing polypeptide, wherein said first human CH3-containing polypeptide comprises a replacement of the amino acid at position 392 with a negative-charged amino acid and said second human IgG CH3-containing polypeptide comprises a replacement of Asp399, Glu356, Asp356, or Glu357 with a positive-charged amino acid. 
     
     
         2 . The host cell of  claim 1 , wherein Lys392 is replaced with a negative-charged amino acid. 
     
     
         3 . The host cell of  claim 1 , wherein Asn392 is replaced with a negative-charged amino acid. 
     
     
         4 . The host cell of  claim 1 , wherein said first human CH3-containing polypeptide further comprises Lys409 or Arg409 replaced with a negative-charged amino acid. 
     
     
         5 . The host cell of  claim 1 , wherein Lys392 or Asn392 is replaced with aspartic acid. 
     
     
         6 . The host cell of  claim 4 , wherein said Lys409 or Arg409 is replaced with aspartic acid. 
     
     
         7 . The host cell of  claim 1 , wherein said second human IgG CH3-containing polypeptide comprises a replacement of Asp399, Glu356, Åsp356, or Glu357 with lysine. 
     
     
         8 . The host cell of  claim 1 , wherein said second human IgG CH3-containing polypeptide comprises a replacement of Asp399 and Glu356 with lysine. 
     
     
         9 . The host cell of  claim 1 , wherein the heterodimeric protein comprises a human IgG Fc region. 
     
     
         10 . The host cell of  claim 9 , wherein the human IgG Fc region comprises an IgG1 Fc region. 
     
     
         11 . The host cell of  claim 9 , wherein the IgG Fc region comprises an IgG2 Fc region. 
     
     
         12 . The host cell of  claim 9 , wherein the IgG Fc region comprises an IgG3 Fc region. 
     
     
         13 . The host cell of  claim 9 , wherein the IgG Fc region comprises an IgG4 Fc region. 
     
     
         14 . The host cell of  claim 1 , wherein the first CH3-containing polypeptide is an antibody heavy chain. 
     
     
         15 . The host cell of  claim 1 , wherein the second CH3-containing polypeptide is an antibody heavy chain. 
     
     
         16 . The host cell of  claim 1 , wherein the heterodimeric protein further comprises one or more antibody light chains. 
     
     
         17 . The host cell of  claim 1 , wherein the heterodimeric protein is selected from the group consisting of an antibody, a bispecific antibody, a monospecific monovalent antibody, a bispecific maxibody, a monobody, a peptibody, a bispecific peptibody, a monovalent peptibody, and a receptor fusion protein. 
     
     
         18 . The host cell of  claim 1 , wherein the host cell is a mammalian host cell. 
     
     
         19 . The host cell of  claim 18 , wherein the mammalian host cell is a Chinese hamster ovary (CHO) cell line. 
     
     
         20 . A host cell for producing a heterodimeric protein, said host cell comprising a nucleic acid encoding a first CH3-containing polypeptide and a nucleic acid encoding a second CH3-containing polypeptide, wherein said first CH3-containing polypeptide comprises replacement of the amino acids at positions 392 and 409 with a negative-charged amino acid and said second CH3-containing polypeptide comprises replacement of the amino acids at positions 356 and 399 with a positive-charged amino acid. 
     
     
         21 . The host cell of  claim 20 , wherein the negative charged amino acid is aspartic acid. 
     
     
         22 . The host cell of  claim 20 , wherein the positive charged amino acid is lysine. 
     
     
         23 . The host cell of  claim 21 , wherein the positive charged amino acid is lysine. 
     
     
         24 . The host cell of  claim 20 , wherein the mammalian host cell is a Chinese hamster ovary (CHO) cell line. 
     
     
         25 . A host cell for producing a heterodimeric protein, said host cell comprising a nucleic acid encoding a first CH3-containing polypeptide and a nucleic acid encoding a second CH3-containing polypeptide, wherein the first CH3-containing polypeptide comprises replacement of the amino acids at positions 370, 392, and 409 with a negative-charged amino acid and said second CH3-containing polypeptide comprises replacement of the amino acids at positions 356, 357, and 399 with a positive-charged amino acid. 
     
     
         26 . The host cell of  claim 25 , wherein the negative charged amino acid is aspartic acid. 
     
     
         27 . The host cell of  claim 25 , wherein the positive charged amino acid is lysine. 
     
     
         28 . The host cell of  claim 26 , wherein the positive charged amino acid is lysine. 
     
     
         29 . The host cell of  claim 25 , wherein the mammalian host cell is a Chinese hamster ovary (CHO) cell line.