ABSTRACT
A process for large-scale purification of staphylococcal enterotoxin B (SEB) is presented. The process consists of three chromatographic steps. The first two steps are based on ion-exchange chromatography and the final one on gel filtration. Diluted culture supernatant (2000 dm3) with a SEB content of 0.4 mg dm-3 were processed and purified. SEB was obtained in 1.5 dm3 of 0.1 mol dm-3 ammonium hydrogen carbonate buffer, pH 8.0. The final product produced a single band on SDS polyacrylamide gel electrophoresis and a single peak when subjected to analytical gel filtration. The overall recovery was 74%.
Subject(s)
Enterotoxins/isolation & purification , Staphylococcus aureus/growth & development , Chromatography, Gel/methods , Chromatography, Ion Exchange/methods , Electrophoresis, Polyacrylamide Gel/methods , FermentationABSTRACT
Albumin has been purified by chromatography using standardized procedures with different starting materials depending on previous fractionation. Plasma can be used directly after cryoprecipitation and Factor IX adsorption or after isolation of IgG. The plasma was centrifuged and then desalted on Sephadex G-25 Coarse, the pH was adjusted, and the euglobulins were precipitated before ion-exchange chromatography on DEAE- and CM-Sepharose CL-6B. This was followed by concentration by ultrafiltration, gel filtration on Sephacryl S-200, and final concentration and formulation. The albumin obtained was 99% pure and contained less than 1% of aggregated protein.
