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The Journal of Immunology, 1968, 100, 942 -954
Copyright © 1968 by The American Association of Immunologists, Inc.
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Equine Antibodies to Human {gamma}G-Globulin1

II. Isolation and Antigenic Analysis of {gamma}2- and {gamma}1-Antibody Fractions from Equine Antisera to Human {gamma}G-Globulin

Sarah L. Johnston and Peter Z. Allen

From the Department of Microbiology, University of Rochester School of Medicine and Dentistry, Rochester, New York 14620

Abstract

Successive bleedings were obtained from a single horse during the course of immunization with human {gamma}G-globulin. Each antiserum was separated into {gamma}1- and {gamma}2-antibody fractions by a procedure involving salt fractionation followed by sequential anion and cation exchange chromatography. Isolated {gamma}1-antibody fractions from each bleeding gave a flocculating type quantitative precipitin curve while isolated {gamma}2-antibody fractions gave the conventional type precipitin curve. Solubilities of specific precipitates obtained with {gamma}1 and {gamma}2 fractions were estimated from the slopes of volume dependence curves and specific solubilities of 2 to 3 and 14 to 18 µg total N/ml were found for {gamma}2 and {gamma}1 specific precipitates, respectively.

Specific precipitates solubilized by excess antigen were examined with antisera to equine serum components. Each solubilized {gamma}2 specific precipitate was shown to contain {gamma}Ga- and/or {gamma}Gb-globulins. Solubilized specific precipitates prepared from isolated {gamma}1 fractions of each bleeding were shown to contain at least four components. Two components could be identified as IgG(T) and IgG immunoglobulins while a third component having a mobility between that of {gamma}2- and {gamma}1-globulin and a fourth with {gamma}1 mobility were not identified and remain to be classified.

Footnotes

This study was supported by United States Public Health Service Research Grant A1-06534. Presented in part to the American Association of Immunologists, Chicago, Illinois, April 1967.






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