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On the Failure of Concentrated Diphtheria Antitoxin to Give the Ramon Flocculation Reaction with Toxin

From the Department of Bacteriology, School of Medicine, University of Pennsylvania, Philadelphia

Abstract

No satisfactory explanation has yet been offered for the fact that highly refined and concentrated solutions of pseudoglobulin prepared from diphtheria antitoxin frequently do not give the Ramon flocculation reaction with toxin, despite the fact that their biological activity as measured by protection experiments on guinea pigs may greatly exceed that of the original serum.

The following experiments were undertaken in order to ascertain whether the absence of flocculation is due to the failure of the antitoxin globulin to combine with toxin in vitro.

Varying quantities of anti-diphtheria horse serum pseudoglobulin were added to one L+ dose of toxin, the mixture was allowed to stand for one-half hour, and the globulin was then precipitated by the addition of a rabbit anti-horse serum. The latter was added until the supernatant fluid contained demonstrable excess of the anti-horse serum antibody, and no demonstrable horse serum protein. The mixture was then centrifuged, and the supernatant fluid tested for its toxin content.