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The Quantitative Precipitin Reaction of Equine Diphtheria Antitoxin

From the Biologic Laboratories, Institute of Laboratories, Massachusetts Department of Public Health, and the Department of Bacteriology and Immunology, Harvard Medical School Boston, Mass.

Abstract

Pappenheimer and Robinson (1) studied the quantitative reaction between diphtheria antitoxin (equine) and toxin. Their curves showed inhibition of precipitation in excess antibody and excess antigen and a wide zone of equivalence as determined by the rabbit intracutaneous test. The points in the equivalence zone defined a straight line whose slope they interpreted as a measure of specifically precipitable antigen nitrogen (N) and whose intercept measured specifically precipitable antitoxin N. They concluded that co-precipitation of non-toxin material did not occur in appreciable amounts.

This interpretation has been seriously criticized by Pope and his co-workers (2), who have been unable, in general, to reproduce the curve as originally described by Pappenheimer and Robinson. Pope concludes, after extensive experimentation, that co-precipitation of other antigen-antibody complexes definitely contributes to the total N. Indeed, he suggests that co-precipitates may actually represent a major part of the total precipitate with the true toxin-antitoxin complex precipitating, quite incidentally, simultaneously with other immunologic complexes present in the mixture.