Production of heteromultimeric proteins

Abstract

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.

What is claimed is: 
     
         1 . A method of preparing a heteromultimeric protein comprising a first hinge-containing polypeptide having a first heterodimerization domain and a second hinge-containing polypeptide having a second heterodimerization domain, wherein the second heterodimerization domain interacts with the first heterodimerization domain, and wherein the first and second hinge-containing polypeptides 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 hinge-containing polypeptide under conditions where the hinge-containing polypeptide is expressed;   (b) culturing a second host cell comprising a nucleic acid encoding the second hinge-containing polypeptide under conditions where the hinge-containing polypeptide is expressed;   (c) disrupting the cell membranes of the first and second host cells, wherein the first and second host cells have been combined together in a single suspension; and   (d) recovering the heteromultimeric protein,   
       wherein said method does not require the addition of a reductant. 
     
     
         2 - 89 . (canceled)