Purification by high performance liquid chromatography of Clostridium perfringens type A enterotoxin prepared from high toxin producers selected by a toxin-antitoxin halo.

摘要: High enterotoxin-producing substrains of Clostridium perfringens type A were selected reproducibly as colonies having toxin-antitoxin haloes on agar plates Duncan-Strong medium containing antitoxin serum. Enterotoxin from these sub strains was subjected to rapid purification by high performance liquid chromatography (HPLC). For this, the toxin extracted sonication sporulating bacteria grown in sporulation medium, fractionated ammonium sulfate (40% saturation) precipitation and differential solubilization then purified HPLC: gel permeation through a G2000SW column ion-exchange Mono Q column. Purified preparations had similar specific activity (4.2 x 102 mouse MLD/mg protein) homogeneity polyacrylamide gel-electrophoresis obtained conventional Sephadex-G200 By further HPLC column, minor nontoxin proteins separated without loss toxicity protein basis. The final yield about 15° that bacterial extract. two procedures each took only one hour.