title: Detection, isolation and characterization of staphylococcal enterotoxin B in protein A preparations purified by immunoglobulin G affinity chromatography.
abstract: Studies were performed to detect and isolate trace contaminants of staphylococcal enterotoxin B (SEB) in various protein A preparations isolated by affinity chromatography employing human IgG covalently bound to Sepharose 4B. Utilizing an ELISA technique, trace amounts (0.018-0.138%) of SEB could be detected in protein A preparations after separation of the SEB employing a molecular sizing column in a high pressure liquid chromatography (HPLC) system. Trace contamination by SEB could be removed from protein A preparations by an additional DEAE ion exchange chromatography step employing a low ionic strength buffer system (0.005 M NaCl in 0.01 phosphate buffer, pH 7.50). The resulting protein A preparations possessed a purity higher than that observed prior to the final purification step. Polyacrylamide gel electrophoresis (PAGE) analyses of the trace contamination removed from protein A preparations by ion exchange chromatography revealed, in addition to SEB, several additional contaminating polypeptides of an unknown nature. These studies indicate that protein A preparations of high purity can be prepared by employing DEAE ion exchange chromatography in addition to affinity chromatography utilizing immobilized human IgG.
authors.affiliation: IMRE Corporation Seattle, WA 98109.
authors.forename: J
authors.initials: J
authors.lastname: Balint
authors.forename: C
authors.initials: C
authors.lastname: Totorica
authors.forename: J
authors.initials: J
authors.lastname: Stewart
authors.forename: S
authors.initials: S
authors.lastname: Cochran
mesh_terms: D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004768:Enterotoxins / Q000032:analysis*; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A / Q000032:analysis*
chemical_list: D004768:Enterotoxins; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A; C031818:enterotoxin B, staphylococcal