Effect of single peptide bond scission by trypsin on the structure and activity of staphylococcal enterotoxin B.
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Biomedical subjects
Publications and source records attributed to J F Metzger.
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Release of Staphylococcus aureus enterotoxin B (SEB) into the culture medium was initiated during the mid-log phase of growth. A medium consisting of 4% N-Z Amine A (Sheffield), 0.2% dextrose, and 1% yeast extract supported maximum production of SEB. Although pH of the medium during cultivation did not significantly affect the growth curve of the organism, the time required for detectable excretion was affected, as was the final yield. Optimal conditions for SEB production were achieved with pH control at 7.0; alkaline control (pH 8.0) produced only minimal amounts of toxin, whereas acid control (pH 6.0) resulted in 50% reduction in yield. Slightly less SEB was produced when there was no extrinsic pH control, and cultures were buffered only by media constituents and by-products of growth. With pH control at 7.0, deletion of 0.2% dextrose from the medium resulted in 40% reduction in the 8-h yield. There was also a delay in production during early stages of fermentation.
A rapid solid-phase radioimmunoassay for staphylococcal enterotoxin A is described. The assay procedure requires 3 to 4 h for completion by using a competitive inhibition system in which the antibody is attached to bromacetyl cellulose particles. It is accurate to a level of 0.01 mug of enterotoxin A/ml in a variety of media such as ham, milk products, crab meat, custard, etc. No significant interference was found with any media or food product tested.
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The need for a human-derived immune globulin to replace the equine antitoxins currently used in the treatment of botulism is well recognized. A small group of individuals who had received multiple immunizations with pentavalent botulinal toxoid were plasmapheresed for the purpose of collecting a botulism-immune plasma of human origin to be fractionated for the production of immune globulin. Human-derived immune globulin will offer the advantage over equine antitoxins of not inducing reactions to foreign protein and of having a prolonged effective half-life.