Preparation of labeled staphylococcal enterotoxin A with high specific activity

Abstract

Staphylococcal enterotoxin A (SEA) was labeled by the chloramine-T method with 125I to a specific activity of 68 to 300 muCi per mug of SEA and with 131I to specific activity of 8 to 218 muCi per mug of SEA. SEA was partially damaged and aggregated during the labeling and storage. The damage seemed not to be greatly dependent on the specific activity of labeled entertoxin. Crossed immunoelectrophoresis showed two antigenically active and three inactive components in the ascending part of the labeled enterotoxin peak during fractionation by gel chromatography. During storage at 4 degrees C, the antigenic activity of label decreased faster when labeling had been with 131I than when with 125I. The antigenic activity of labeled SEA was lowered remarkably in the ascending part of the protein peak. Greatest release of radioiodine during storage was in the same part of protein peak. According to these results, the most suitable label for radioimmunoassay is obtained from the descending part of protein peak.