A high-performance liquid chromatographic procedure for the purification of mouse monoclonal antibodies

Abstract

High-performance liquid chromatography was applied to the purification of monoclonal antibodies from mouse ascites fluid. The method was based on anion-exchange chromatography using a TSK DEAE-5PW column and a gradient elution with 20 mM Tris, pH 8.5, and 20 mM Tris, pH 8.5, containing 2.0 M sodium acetate. The method can be applied to analytic or preparative scale separations. Purified immunoglobulins were isolated from samples of 20 to 100 microliter containing up to 19 mg total protein. The average recovery of total protein was 89 +/- 12%. Recovery of the immunoglobulins, based on recovery of immunological activity, was quantitative. In addition to separating the immunoglobulins from the other serum proteins, the various classes of IgG were resolved.