Isolation and purification of Clostridium perfringens enterotoxin by affinity chromatography.

摘要: Sporulating cells of certain strains Clostridium perfringens produce an intracellular enterotoxin believed to be the causative agent C. food poisoning. Rabbit antibody preparations both vegetative and sporulating cell extracts were used purify toxin by differential immuno-affinity chromatography. Solid phase immune adsorbents prepared attaching via α-amino groups succinylaminoethyl Sepharose-4B give two resins: one binding anti-vegetative immunoglobulin G (IgG), (V-resin), other, antisporulating IgG, (S-resin). extract was passed through resin with bound IgG extract. A large protein fraction representing antigens common forms retained, but not measurably absorbed as determined erythemal activity in rabbits. The toxic then majority did adhere this resin; however, that which did, showed Disc-gel electrophoresis eluted from demonstrated presence five components. One elicited maximum capacity V-resin column 102 μg protein/ml resin, S-resin column, 32 resin. 150-fold purification achieved procedure. Resins could repeatedly.