Patent ID: 9670269
Date: 2017-06-06
CPC Classifications: A61P,C07K

Claim:
1. A method for producing a modified multispecific human or humanized antibody comprising a first polypeptide and a second polypeptide, wherein the method comprises the steps of: (a) providing a pair of nucleic acids respectively encoding a pair of polypeptides, wherein the pair of polypeptides is either a pair of human or humanized antibody heavy chains or a pair of human or humanized antibody light chains; (b) modifying one or both of the nucleic acids to alter one or more surface amino acid residues of the encoded polypeptide(s), thereby generating a modified pair of nucleic acids encoding a modified pair of polypeptides, wherein the modification increases the difference between the respective isoelectric points of the pair of polypeptides; (c) providing host cells containing the modified pair of nucleic acids; (d) culturing the host cells so that they express the modified pair of nucleic acids; and (e) using a method comprising ion exchange chromatography to purify, from the host cell culture, a modified human or humanized multispecific antibody comprising the modified pair of polypeptides, wherein ion exchange chromatography separates the modified human or humanized multispecific antibody from homomultimers formed from each polypeptide of the modified pair of polypeptides based on the difference between the respective isoelectric points of the modified pair of polypeptides, wherein the one or more surface amino acid residues that are altered comprise at least one amino acid residue located at a position or positions selected from heavy chain variable region positions 1, 3, 5, 8, 10, 12, 13, 15, 16, 19, 23, 25, 26, 27, 39, 42, 43, 44, 46, 68, 71, 72, 73, 75, 76, 81, 82b, 83, 85, 86, 97, 105, 108, 110, and 112 (Kabat numbering), heavy chain constant region positions 137, 196, 203, 214, 217, 233, 268, 274, 276, 297, 355, 392, 419, and 435 (EU numbering), and light chain variable region positions 1, 3, 7, 8, 9, 11, 12, 16, 17, 18, 20, 22, 38, 39, 41, 42, 43, 45, 46, 49, 57, 60, 63, 65, 66, 68, 69, 70, 74, 76, 77, 79, 80, 81, 85, 100, 103, 105, 106, 107, and 108 (Kabat numbering).