Immunoselection of B cell hybridomas with anti-idiotypic antisera to study the role of heavy and light chains for idiotype expression and antibody activity.
Conventional reconstitution experiments with heavy (H) and light (L) offins of hybridoma anti-poly-(Glu60Ala30Tyr10) (anti-GAT) antibodies demonstrated that H and L chains are required for efficient expression of anti-GAT activity and the previously defined common CGAT idiotype. On the basis of this observation, we treated a CGAT+ hybridoma cell line (F9.195.6), which was produced by fusion with an MOPC 21 (gamma 1, K) secretor line, with anti-CGAT idiotypic antibodies plus complement to select CGAT-variants. Such treatment greatly enhanced the frequency of isolating hybridoma variants. Molecules secreted from these variants bear anti-GAT mu-chain and MOPC 21 L chain (as judged by serology and Southern blot studies), lack GAT binding activity and CGAT idiotype, and exhibit a "new" private idiotype (E8). Anti-private idiotypic sera were successfully used to select a series of CGAT+, E8- clones that secrete molecules bearing only the appropriate anti-GAT mu- and K chains. From E8- hybridoma supernatants, CGAT- molecules bearing MOPC 21 H chain and anti-GAT L chain could be obtained. The data indicate that immunoselection of hybridoma clones with anti-idiotypic antibodies is a convenient means of preparing reconstituted antibody molecules. Furthermore, the data demonstrate that CGAT idiotype determinants are expressed on antibody molecules composed of appropriate H and L chains.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000502:physiology*; D000961:Antilymphocyte Serum / Q000276:immunology / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000276:immunology*; D001666:Binding Sites, Antibody; D014644:Genetic Variation; D006825:Hybridomas / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000502:physiology; D007130:Immunoglobulin Idiotypes / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000502:physiology; D007135:Immunoglobulin Variable Region; D051379:Mice; D008811:Mice, Inbred DBA; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D011108:Polymers
D000888:Antibodies, Anti-Idiotypic; D000961:Antilymphocyte Serum; D007143:Immunoglobulin Heavy Chains; D007130:Immunoglobulin Idiotypes; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D010455:Peptides; D011108:Polymers; C020749:GAT