Production of heteromultimeric proteins

摘要

Described herein are methods for the efficient production of antibodies and other multimeric protein complexes (collectively referred to herein as heteromultimeric proteins) capable of specifically binding to more than one target. The targets may be, for example, different epitopes on a single molecule or located on different molecules. The methods combine efficient, high gene expression level, appropriate assembly, and ease of purification for the heteromultimeric proteins. The invention also provides methods of using these heteromultimeric proteins, and compositions, kits and articles of manufacture comprising these antibodies.

主权项

1. A method of preparing a heteromultimeric protein comprising a first antibody heavy chain paired with a first antibody light chain and a second antibody heavy chain paired with a second antibody light chain, and wherein the first and second antibody heavy chains are linked by at least one interchain disulfide bond, the method comprising the steps of:
(a) culturing a first host cell comprising a first nucleic acid encoding the first antibody heavy chain and the first antibody light chain under conditions where a first half antibody comprising the first antibody heavy chain and the first antibody light chain is expressed; (b) culturing a second host cell comprising a second nucleic acid encoding the second antibody heavy chain and the first antibody light chain under conditions where a second half antibody comprising the second antibody heavy chain and the second antibody light chain is expressed; (c) combining cultures of the first and second host cells to produce a combined culture comprising the first host cells and second host cells, wherein the heteromultimeric protein is formed in the combined culture; and (d) recovering the heteromultimeric protein without prior recovery or purification of the first and second half antibodies from separate cultures.