Purification of staphylococcal enterotoxins B and C-2 by dye ligand chromatography production of antisera

Enterotoxins B and C-2 were purified from sac culture supernatants by a combination of Amberlite CG-50 (cation exchange) chromatography and Red A ligand chromatography. One-hundred milliliters of initial supernatant fluid containing 270 mu g per milliliter of SEB and 150 ml of supernatant fluid containing 93 mu g per milliliter of SEC(2) gave overall yields of 59% and 42%, respectively. The purified toxins were homogeneous when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Rabbits were immunized with SEB and SEC(2). Maximal titers were 80 for SEB and 40 SEC(2). The methodology described herein can be employed to purify SEB and SEC(2) for use as reagents and as potent immunogens by processing low initial volumes of cultures supernatants.