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A microELISA useful for determination of protein A-binding monoclonal antibodies
1Department of Medical and Physiological Chemistry, University of Uppsala, Sweden.
Affinity chromatography on protein A columns is widely used for the purification of monoclonal antibodies. However, some monoclonal antibodies do not bind protein A. We have developed a sandwich-ELISA for determination of the protein A binding capacity of antibodies. This utilizes microtitre plates coated with protein A, and labeled chicken antibodies for detection of bound antibody. Chicken IgG is one of the few antibodies that does not react with protein A, and can thus be used without the problem of a high background.
Affinity chromatography on protein A columns is widely used for the purification of monoclonal antibodies. However, some monoclonal antibodies do not bind protein A. We have developed a sandwich-ELISA for determination of the protein A binding capacity of antibodies. This utilizes microtitre plates coated with protein A, and labeled chicken antibodies for detection of bound antibody. Chicken IgG is one of the few antibodies that does not react with protein A, and can thus be used without the problem of a high background.