Characterization of molecular heterogeneity and multispecificity in homologous idiotypic antisera.
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 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.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000302:isolation & purification; D000918:Antibody Specificity; D000939:Epitopes; D005260:Female; D006386:Hemagglutination Tests; D007135:Immunoglobulin Variable Region; D007136:Immunoglobulins; D007525:Isoelectric Focusing; D051379:Mice; D008805:Mice, Inbred A; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D009194:Myeloma Proteins / Q000276:immunology; D011863:Radioimmunoassay
D000888:Antibodies, Anti-Idiotypic; D000939:Epitopes; D007135:Immunoglobulin Variable Region; D007136:Immunoglobulins; D009194:Myeloma Proteins