Large amounts of high-titre α-haemolysin were produced in a nutrient broth medium. Production was enhanced by growth in a medium containing large molecular-weight components. The haemolysin was purified by “salting out” with ammonium sulphate precipitation at its iso-electric point 4·6) and by chromatography on Sephadex G-200. Evidence is presented that the haemolysin is a large protein (approximate molecular weight = 580,000) that was not easily dissociated into smaller subunits. Lethality in mice and rabbits was not observed after intravenous injection of α-haemolysin, but hard swellings appeared at injection sites after the intradermal administration of active haemolysin preparations. The haemolysin did not cause fluid accumulation in ligated intestinal segments in rabbits.


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