Purification of in vitro produced mouse monoclonal antibodies. A two-step procedure utilizing cation exchange chromatography and gel filtration.

A method for production of purified and concentrated mouse monoclonal antibodies was developed. The rationale for the procedure is, firstly, to expand hybridoma cell number in culture medium-containing serum; secondly, to transfer the cells to serum-free medium for production of antibodies; and finally, to harvest antibodies from the conditioned medium by means of cation exchange chromatography on SP-Sephadex C-50, followed by gel filtration on Superose 6B. When compared with chromatography of monoclonal antibodies on protein A-Sepharose our results suggest that the method is particularly useful for purification of antibodies of the IgG1 subclass. Experience from production and purification of 15 various monoclonal antibodies is reported.