Purification and Characterization of Tetanus Toxoid and Toxin

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Abstract
Summary: Filtration of partially purified tetanus toxoid through the dextran gel Sephadex G-100 has yielded at least four clearly identifiable fractions. The two fractions emerging first from the column, containing 55 to 65% of the nondialyzable nitrogen in the starting material, were capable of reacting in vitro with tetanus antitoxin and stimulating immunity to challenge with tetanus toxin in mice. The antigenicity of these fractions did not differ significantly from the parent toxoid. The third fraction emerging from the column, which apparently is of smaller molecular size, reacted in vitro with tetanus antitoxin, but was poorly antigenic in animals. The fourth fraction identified, presumably consisting of nondialyzable medium derivatives and nonspecific by-products of metabolism of the tetanus organism, was inactive both in vitro and in vivo. The results of this study have demonstrated the value of gel filtration for the analysis of tetanus toxoid as well as for its purification. The simplicity of the technique as well as the low cost of materials are additional advantages of the system. The method described has yielded satisfactory separation when applied to batches of toxoid containing upwards of 200,000 flocculation units.

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