Method for making Heteromultimeric molecules

摘要

Methods for making heteromultimeric molecules, such as bispecific antibodies, and compositions comprising these molecules are disclosed. The methods include introducing mutations in amino acids that are in contact at the interface of two polypeptides, such that the electrostatic interaction between the ion pairs is altered.

主权项

1. A method of preparing a bispecific antibody that specifically binds VEGF and DLL4, wherein the bispecific antibody comprises a first and a second human immunoglobulin heavy chain polypeptide and a first and a second immunoglobulin light chain polypeptide, wherein the first and second immunoglobulin heavy chain polypeptide each comprise a CH3 domain and the CH3 domains comprise substitutions that promote heterodimerization of the first and second immunoglobulin heavy chain polypeptides, and wherein the first and second light chain polypeptides are identical in amino acid sequence, the method comprising substituting amino acids residues within the immunoglobulin heavy chain CH3 domains, and wherein the bispecific antibody is selected from the group consisting of:
(a) a human IgG1, wherein the substitutions consist of: a glutamate or aspartate at positions in the first immunoglobulin heavy chain polypeptide corresponding to positions 253 and 292 of SEQ ID NO:33, and a lysine at positions in the second immunoglobulin heavy chain polypeptide corresponding to positions 240 and 282 of SEQ ID NO:33; (b) a human IgG2, wherein the substitutions consist of: a glutamate or aspartate at positions in the first immunoglobulin heavy chain polypeptide corresponding to positions 249 and 288 of SEQ ID NO:34 and a lysine at positions in the second immunoglobulin heavy chain polypeptide corresponding to positions 236 and 278 of SEQ ID NO:34; (c) a human IgG3, wherein the substitutions consist of: a glutamate or aspartate at positions in the first immunoglobulin heavy chain polypeptide corresponding to positions 300 and 339 of SEQ ID NO:35, and a lysine at positions in the second immunoglobulin heavy chain polypeptide corresponding to positions 287 and 329 of SEQ ID NO:35; and (d) a human IgG4, wherein the substitutions consist of: a glutamate or aspartate at positions in the first immunoglobulin heavy chain polypeptide corresponding to positions 250 and 289 of SEQ ID NO:36, and a lysine at positions in the second immunoglobulin heavy chain polypeptide corresponding to positions 237 and 279 of SEQ ID NO:36.