High-Performance Liquid Affinity Chromatographic Separation of Mouse Monoclonal Antibodies with Protein A Silica

• Protein A, a bacterial cell wall protein found in Staphylococcus aureus, has been widely used for the analysis of immunoglobulins. By attaching protein A to a microparticulate silica support, a rapid and efficient chromatographic sorbent has been created for the separation of monoclonal antibodies. Examples are given of rapid separations (within 10 min) of murine monoclonal antibodies, belonging to various IgG subclasses and including IgGl. The monoclonal antibodies were isolated with a high purity and with 60–90% recovery of activity. The high-performance liquid affinity chromatography technique based on protein A provides a useful method for monitoring monoclonal antibodies in crude samples, such as ascites and cell culture supernatants. 

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