title: Recovery of staphylococcal enterotoxin from foods by affinity chromatography.
abstract: Extraction, concentration, and serological detection of staphylococcal enterotoxins from foods are laborious and time consuming. By exposing food extracts to an insoluble matrix tagged with specific anti-enterotoxin B, we have been able to recover the toxin from foods in a sensitive and rapid way. After mixing the reagents for 2 h at room temperature, immunoglobulin G antibodies were attached to CNBr-activated Sepharose 4B at pH 8.5 (0.1 M carbonate buffer with 0.5 M NaCl). Sepharose-antibody complex (1 ml) specifically recovered 0.1 to 30 mug of enterotoxin B from 400 ml of food extract (100 g of food) after mixing for 2 h at 4 C. The Sepharose-antibody-toxin complex was washed with 0.02 M phosphate-buffered saline at pH 7.2, and the toxin was dissociated by 2 to 4 ml of 0.2 M HCl-glycine plus 0.5 M NaCl buffer at pH 2.8. The recovered enterotoxin was free of interfering food components and could be detected serologically. Work to couple antibodies A, B, C, D, and E to Sepharose to recover all five toxins in one step is under study.
mesh_terms: D000936:Antigen-Antibody Complex; D002846:Chromatography, Affinity; D004768:Enterotoxins / Q000032:analysis*; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D013210:Staphylococcus
chemical_list: D000936:Antigen-Antibody Complex; D004768:Enterotoxins