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Quantitative Determination of Antigen, Antibody and Complement in Precipitates

From the Institute for Biological and Medical Chemistry, University of Istanbul

Abstract

1. Immune sera to arsanil-azo-globulin were precipitated by small and by large amounts of antigen, in the presence of active and of inactivated guinea-pig serum. Gravimetric analysis of the precipitates demonstrated:

a) The weight of the precipitate obtained with active guinea-pig serum is greater. Maximally 0.15 to 0.20 mg of complement-protein were fixed per ml of guinea-pig serum.

b) The amount of complement protein in the precipitate was independent of the antigen-antibody ratio and equaled about 20 per cent of the weight of the precipitates.

c) Neither the antigen-antibody ratio nor the absolute amount of antigen or antibody in the precipitate is altered by complement fixation.

2. Hemolysis of a single red cell requires approximately 10^–11g, i.e. more than 10¹⁰ molecules of saponin or of sodium oleate. The same effect is produced by 1.5 x 10^–14g, i.e. 60000 molecules of complement-protein. The much higher efficiency of complement supports the enzyme theory of complementary action.