Methods and compositions for increasing antibody production

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.