Two enzyme-linked immunosorbent assays were developed for detection of staphylococcal exfoliative toxins A and B (ETA and ETB) with a double-antibody sandwich protocol. Antibodies against both toxins were purified by affinity chromatography from sheep antisera raised against purified ETA and ETB. These affinitypurified antibodies were free of detectable amounts of antibodies to other staphylococcal antigens and neutralized the actions of ETA and ETB. Alkaline phosphatase was conjugated to these antibodies. The enzyme-linked immunosorbent assay, which could detect at least 3 ng of ETA and ETB per ml, was used to quantitate the toxins in the culture supernatant fluids of staphylococcal strains. Thus, the kinetics of ETA and ETB synthesis and of ETA and ETB release into the supernatant fluids were determined; other determinations included the roles of carbon dioxide concentration, pH, glucose concentration, temperature, and agitation on the production of ETA and ETB.