Characterization of the heavy and light chain immunoglobulin variable region genes used in a set of anti-digoxin antibodies.
Five murine A/J hybridomas (the 35-20 group) produce anti-digoxin antibodies that have homologous heavy and light chain immunoglobulin variable regions (VH and VL), yet differ from each other in fine specificity and affinity for digoxin and related cardiac glycosides [Mudgett-Hunter et al. Molec. Immun. 22, 477-488 (1985)]. To determine the origin of the VH and VL genes used in this set of hybridomas, the rearranged VH and VL genes from one of the 35-20 group hybridomas, 40-140, were cloned. The expressed V region, the leader exon and the 5' transcription control regions were sequenced. VH40-140 is a member of the VH36-60 gene family and has greater than 90% homology with several members of that family. A VH40-140 hybridization probe detected two members of the VH36-60 gene family not previously described. The VL40-140 region, a member of the Vk9 subgroup, is nearly identical to the Vk region used by the myeloma T1. Southern analysis with several restriction endonucleases and hybridization probes indicated that the 35-20 group hybridomas each use the same heavy and light chain variable region gene segments in the assembly of their expressed antibody genes. Functional rearranged antibody genes were detected with JH and VH heavy chain probes and with Jk and Vk light chain probes. The availability of the clones of the one heavy and the one light chain variable region gene segment used by the 35-20 hybridoma group will facilitate the use of in vitro mutagenesis in studies of the structural basis of fine specificity in the digoxin antigen-antibody system.
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Cellular and Molecular Research Laboratory, Massachusetts General Hospital, Boston.
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D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular; D004077:Digoxin / Q000235:genetics* / Q000276:immunology; D005796:Genes; D006825:Hybridomas; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D015183:Restriction Mapping
D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D004077:Digoxin