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Sepharose gels as antigenic matrices for specific binding of anti-galactosyl antibodies
D Hädge1, I Podszuck, H Ambrosius
1Laboratory of Immunobiology, Karl Marx University Leipzig, GDR.
From 100 ml of pooled normal serum taken from different species (man, cow, rabbit, chicken, duck, carp) 0.5 to 4.0 mg of high and low molecular weight anti-galactosyl antibodies could be isolated by affinity-chromatography on non-ligand-attached Sepharose 4B. The antibodies bound to the agarose matrix were eluted with 0.5 M D-galactose containing buffers. Using species-specific anti-immunoglobulin sera in immunoelectrophoresis, the immunoglobulin character of the protein fractions prepared was analyzed. In addition, the H and L chain structure of the anti-galactosyl antibodies was checked by SDS-PAGE.
From 100 ml of pooled normal serum taken from different species (man, cow, rabbit, chicken, duck, carp) 0.5 to 4.0 mg of high and low molecular weight anti-galactosyl antibodies could be isolated by affinity-chromatography on non-ligand-attached Sepharose 4B. The antibodies bound to the agarose matrix were eluted with 0.5 M D-galactose containing buffers. Using species-specific anti-immunoglobulin sera in immunoelectrophoresis, the immunoglobulin character of the protein fractions prepared was analyzed. In addition, the H and L chain structure of the anti-galactosyl antibodies was checked by SDS-PAGE.