Abstract
Injection of a wide variety of animal species with homologous or heterologous myelin basic protein (BP) emulsified with complete Freund's adjuvant (CFA) produces the autoimmune demyelinating disease, experimental allergic encephalomyelitis (EAE). As a first step in the study of antigenic determinants of bovine BP (BBP), specific antibodies were purified from rabbit serum by adsorption on Sepharose-BBP affinity columns and elution with 0.1 M acetic acid. Yields of 0.02–0.50 mg protein per milliliter of serum were obtained from the acetic acid eluates. Immunoelectrophoresis and polyacrylamide gel electrophoresis indicated that the acid eluted material was IgG. The acid eluted IgG formed stable precipitin bands with BBP in agarose gels and also demonstrated anti-BBP activity in a sodium sulfate precipitation radioimmunoassay and in a solid phase radioimmunoassay. Quantitative precipitation of purified antibodies was used to estimate the number of antigenic determinants in the BBP molecule. From a plot of the antibody to antigen ratio of precipitates vs the log of antigen in the precipitates at antibody excess, an effective antigenic valence of approximately four was indicated.
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