Bio-Rad Laboratories, Inc.

Gary S Ott

Hector Juarez-Salinas

Sheldon C Engelhorn

A novel method is disclosed for the purification of monoclonal antibodies from antibodies of the same isotype but different light chain composition which are frequently present in the fluids in which such antibodies are generated. According to the method, the mixture is placed in a hydroxylapatite chromatography column and eluted with a buffer solution in a concentration gradient elution. With the appropriate optimization of system parameters, a separation of antibodies effective for both analytical and preparative purposes is obtained.

Use of hydroxylapatite for the analysis and preparation of purified

Unusually strong binding of IgG to protein A is achieved by contacting these components in the presence of a medium containing a high concentration of salt. Although such binding is of general utility, a particularly useful application is the purification of monoclonal antibodies from ascites fluid by affinity chromatography.

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Livermore, CA, United States of America

Gary S Ott