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Characterization of Molecular Heterogeneity and Multispecificity in Homologous Idiotypic Antisera¹

From the Department of Microbiology and Immunology, and Department of Pathology, Washington University School of Medicine, Division of Biology and Biomedical Sciences, St. Louis, Missouri 63110

Abstract

The molecular heterogeneity of homologous anti-idiotypic reagents was characterized by a novel isoelectric focusing procedure. Idiotypic antisera directed against the PC-binding plasmacytoma protein T15 were raised in CE and A/J mice. These antisera were shown to be highly specific by hemagglutination with myeloma protein-derivatized sheep erythrocytes and by radioimmunoassay. Competition experiments performed with affinity-labeled T15 revealed that about 40% of the pooled CE antibody activity was directed against binding site-associated determinants. Further analysis of anti-idiotypic sera from individual animals with the use of isoelectric focusing disclosed heterogeneous populations of antibody molecules distinguishable by isoelectric point and by subspecificity. Each animal expressed a unique spectrotypic profile. In addition, clones reactive with binding site and non-binding-site determinants as well as some clones with specificity for other PC-binding mouse myeloma proteins were detected. These results emphasize the importance of careful selection and thorough absorption of idiotypic antisera.

Footnotes

¹ This work was supported by United States Public Health Service Grants Al-11635 and CA-16032, National Science Foundation grant PCM76-09719, and by the following companies: Brown & Williamson Tobacco Corporation; Larus and Brother Company, Inc; Liggett & Myers, Incorporated; Lorillard, a Division of Loews Theatres, Inc.; Philip Morris, Inc.; R. J. Reynolds Tobacco Company; United States Tobacco Company; and Tobacco Associates, Inc.

² Supported by the Medical Scientist Training Program Grant GM-02016.