Routine large-scale production of monoclonal antibodies in a protein-free culture medium

Abstract

A defined culture medium and cultivation technique are described which allow the long-term production on a large scale of monoclonal antibodies and myeloma proteins without the use of serum, serum proteins, or protein-containing liposomes. The high initial purity of the culture supernatants obtained with protein-free medium facilitates the purification of the monoclonal antibodies, the immunoglobulin (Ig) purity of which is limited only by the genetic homogeneity of the cells. Successful growth of 8 hybridoma and 2 myeloma lines has been achieved. SDS-PAGE analysis revealed that the levels of Ig production in the protein-free medium were comparable to those achieved with serum-supplemented media and with media supplemented with purified serum proteins. Stability of Ig production during continuous cultivation in the protein-free medium over an extended period was studied for a myeloma and a hybridoma line, both of which produced high levels (50–200 μg/ml) of Ig for periods of 11 and 22 weeks, respectively.