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Fractionation of Purified Antibodies to the Dinitrophenyl Group with Cross-Reacting Immunoadsorbents¹

From the Department of Microbiology, University of Colorado Medical Center, Denver, Colorado

Abstract

Purified anti-dinitrophenyl group antibodies from pooled antisera or antiserum of a single rabbit were fractionated by means of homologous and cross-reacting immunoadsorbents. Multiple fractions were obtained from all immunoadsorbents by means of stepwise hapten elution with dinitrophenol and when the homologous immunoadsorbent, 2,4-dinitrophenylaminoethyl cellulose, or the extensively altered immunoadsorbent, nitrobenzyl cellulose, were used only a few fractions or a single fraction were obtained. The most effective immunoadsorbent was found to be 2,4,6-trinitrophenylaminoethyl cellulose, which yielded eight major fractions of anti-DNP from the serum of a single rabbit. The chromatographic behavior appears primarily to be determined by differences of the binding ratio of various antibody fractions for the immunoadsorbent and the eluting hapten.

Footnotes

¹ Supported by Grant No. AI 03047 from the National Institute of Allergy and Infectious Diseases, National Institutes of Health. A preliminary report was presented at the 150th National Meeting of the American Chemical Society, September 13 to 17, 1965.