Two fragments with molecular weights of 110,000 and 60,000 were separated in a preparatory scale by gel filtration of the reduced Clostridium botulinum type B trypsinized derivative toxin on Sephadex G-200 with 0.05 M tris(hydroxymethyl)aminomethane-0.38 M glycine buffer, pH 8.3, containing 5 mM ethylenediaminetetraacetate, 1 mM dithiothreitol, and 2 M urea as eluant. They were both antigenic, forming crossing precipitin lines against type B antitoxin in agar gel double diffusion tests.
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