Isolation of Molecules of Restricted Allotype from Antistreptococcal Polysaccharide Antibody

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Abstract
Rabbits were immunized with pepsin-treated Streptococcus pyogenes, strain C-39. Two of thirty-five rabbits immunized responded with antibodies of limited electrophoretic heterogeneity, as indicated by zone electrophoresis and isoelectric focusing. Both of these rabbits were heterozygous, one at the a locus and the other at the b locus. Antibodies in the major peaks of the isoelectric focusing profile were refocused. The antibodies were then distributed in a very narrow range of isoelectric pH. In each case the protein in the peak had a distribution of allotypic specificities markedly different from that of the IgG of the same serum, with one specificity predominating. When light chains were isolated from the preparation heterozygous at the b locus, and subjected to isoelectric focusing, 99% of the chains in the major peak were of b5 specificity. In contrast, isoelectric focusing and refocusing of nonspecific IgG had no appreciable effect on the distribution of allotypic specificities. The results provide support for the hypothesis that a substantial portion of certain antipolysaccharide antibody populations may be of monoclonal origin. The data also suggest that the isoelectric focusing technique may be applicable to the isolation of relatively homogeneous antibody populations on a preparative scale.

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