Patent ID: 7608429

Claim:
A method of producing increased yields of an intact heavy and light chain-comprising antibody, wherein the intact antibody comprises a heavy chain variant of a reference antibody heavy chain, where the variant heavy chain lacks at least one inter-heavy chain hinge region disulfide bond as compared to the reference heavy chain, comprising (a) expressing in a host cell an antibody heavy chain-encoding polynucleotide, and a polynucleotide encoding antibody light chains capable of functionally pairing with the antibody heavy chains encoded by the polynucleotide, wherein the polynucleotide encoding the variant heavy chain polypeptide is made by a method comprising: (i) providing a nucleic acid comprising sequence encoding the reference antibody heavy chain polypeptide having at least one inter-heavy chain hinge region disulfide bond; (ii) modifying the nucleic acid to a variant nucleic acid encoding an antibody heavy chain polypeptide which cannot form at least one of the inter-heavy chain hinge region disulfide bond or bonds formed in the reference heavy chain, wherein the variant hinge region cannot form at least one inter-heavy chain hinge region disulfide bond because it is modified to lack a cysteine residue present in a corresponding non-variant heavy chain polypeptide by either deletion of the cysteine residue or substitution of the cysteine residue with an amino acid residue not capable of forming an inter-heavy chain disulfide bond; (b) culturing the host cell under conditions permissive for expression of the heavy and light chain antibody polypeptides and pairing of the heavy and light chain antibody polypeptides, wherein the amount of intact antibody produced in the host cell is at least 10% greater than the amount of an antibody comprising the corresponding non-variant heavy chain polypeptide expressed under similar culture conditions; and (c) recovering said intact antibody from the host cell.