The Journal of Experimental Medicine
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The Journal of Experimental Medicine, Vol 127, 1003-1011, Copyright © 1968 by The Rockefeller University Press

ARTICLE

AN APPROACH TO THE QUANTITATION OF IMMUNOGENIC ANTIGEN

Patricia J. McConahey 1, Jean-Charles Cerottini M.D.1, and Frank J. Dixon M.D.1

1 From the Department of Experimental Pathology, Scripps Clinic and Research Foundation, La Jolla, California 92037

Using passively administered isotope-labeled anti-KLH to suppress the antibody response of rabbits to KLH, we have attempted to estimate the amount of antigen actually involved in stimulating antibody formation. Single and paired label tracer studies of passively administered anti-KLH IgG indicated that from 0.7 to 2.9 µg were utilized or involved by the antigen in the course of a 90% suppression of the response to 2 mg KLH. Tracer studies of labeled anti-KLH F(ab')2 fragments revealed the retention of from 2 to 3 µg of these fragments in the entire rabbit during a 60% suppression of the response to 1 mg KLH. Based on previously determined ratios of mixtures of KLH and suppressive amounts of anti-KLH in adjuvant, the antibody utilization data were converted to the probable amount of antigen or immunogen involved. It appears that after an injection of 2 mg of KLH approximately 0.2–0.5 µg of antigen persisted and reacted with antibody given 24 hr later. Since all of this persisting, reactive antigen may not be immunogenic, the above estimate of immunogen is probably high, but may serve to establish upper limits for the amounts of immunogen involved in stimulating antibody formation and provide a meaningful frame of reference for antigen tracer studies.

Submitted on January 11, 1968


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