Monoclonal antibodies to human terminal transferase.
Human terminal deoxynucleotidyltransferase was used to produce mouse monoclonal antibodies. Three different assays were used to screen for terminal transferase antibodies and antibody-producing hybridoma cells: an enzyme binding assay, an enzyme-linked immunosorbent assay, and a "terminal transferase-specific antibody" cytochemical procedure that allows visualization of the homogeneity of the hybridoma clones producing antibodies to terminal transferase. Of the 12 mouse hybridoma clones isolated, seven produce IgG1 and five produce IgG2a immunoglobulins. Only one of these monoclonal antibodies is inhibitory to human terminal transferase activity. Four of the monoclonal antibodies react with terminal transferase peptides after separation on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, while the remainder appear to recognize only native determinants on the human enzyme. Cross-reactivity studies with purified calf thymus enzyme show that although four of the monoclonal antibodies bind to native enzyme, none react with calf thymus terminal transferase peptides after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting structural differences within the cross-reactive determinants. A more general survey on the affinity of these monoclonal antibodies to human terminal transferase for the enzyme from other species using an enzyme-linked immunosorbent assay demonstrates that none of the monoclonal antibodies reacts with chicken thymus terminal transferase. The affinity of binding for terminal transferase from other mammals to this panel of monoclonal antibodies varies with the clones as expected, but few bind as strongly as the human enzyme. These results suggest that although antigenic determinants are conserved through evolution, fine structural changes within the determinants do occur.
D000818:Animals; D000911:Antibodies, Monoclonal; D002645:Chickens; D003429:Cross Reactions; D004253:DNA Nucleotidylexotransferase / Q000097:blood* / Q000276:immunology; D004254:DNA Nucleotidyltransferases / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007074:Immunoglobulin G; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000201:enzymology* / Q000276:immunology; D051379:Mice; D013950:Thymus Gland / Q000201:enzymology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D004254:DNA Nucleotidyltransferases; D004253:DNA Nucleotidylexotransferase