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AssigneeChugai Seiyaku Kabushiki Kaisha
InventorsTomoyuki Igawa, Hiroyuki Tsunoda
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The present inventors devised methods for efficiently purifying bispecific antibodies using a chromatography column based on the difference in isoelectric points between the H chains of two types of antibodies, wherein the difference is introduced by modifying the amino acids present on the surface of the antibody variable regions of two types of antibodies that constitute a bispecific antibody. Furthermore, the inventors devised methods for efficiently purifying bispecific antibodies using a chromatography column by linking respective antigen binding sites (heavy chain variable regions) to the antibody constant regions having different isoelectric points, and then coexpressing these antibodies.
A method for producing a multispecific antibody comprising a first polypeptide and a second polypeptide, wherein the method comprises the steps of:
A method for purifying a multispecific antibody comprising a first polypeptide and a second polypeptide, wherein the method comprises the steps of:
A multispecific antibody comprising a first polypeptide and a second polypeptide, wherein the first polypeptide comprises a heavy chain variable region and/or a heavy chain constant region, wherein at least one amino acid residue selected from the amino acid residues at positions 10, 12, 23, 39, 43, and 105, Kabat numbering, in said heavy chain variable region, or the amino acid residues at positions 137, 196, 203, 214, 217, 233, 268, 274, 276, 297, 355, 392, 419, and 435, EU numbering, in said heavy chain constant region, carries a charge, and wherein the isoelectric point of the first polypeptide and that of the second polypeptide are different from each other.
A multispecific antibody in which the isoelectric point of a first polypeptide and that of a second polypeptide are different, and wherein the peaks of the homomultimer of the first polypeptide, the homomultimer of the second polypeptide, and the heteromultimer of the first polypeptide and the second polypeptide are separated in standard chromatography analysis.
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