Production and characterization of monoclonal antibodies against amino-terminus of human alpha-atrial natriuretic polypeptide.
Monoclonal antibodies directed against human, alpha-atrial natriuretic polypeptide (alpha-ANP; Human, 1-28) were obtained by somatic cell fusion between P3-X63-Ag8.653 myeloma cells and spleen cells from a BALB/c mouse immunized with human, alpha-ANP selectively coupled to keyhole limpet hemocyanin. From the analysis of polyclonal sera with respect to determinant specificity before the fusion, the strategy was primarily used to pick up monoclonal antibody specific for the N-terminal residues of human, alpha-ANP. Screening of antibodies in the hybridoma culture supernatants were performed by binding to iodinated synthetic human, alpha-ANP. Two stable clones producing anti-human, alpha-ANP antibodies, designated 13A1 and 10B1, were obtained by the limiting dilution technique. The ability of ANP(Rat, 1-28) to inhibit binding of 125I-human, alpha-ANP to these antibodies was almost equipotent to ANP(human, 1-28). However, ANP fragments (Human, 7-28) and (18-28) did not compete the binding completely. These results suggest that both 13A1 and 10B1 monoclonal antibodies can specifically recognized N-terminus of human, alpha-ANP, and may be a useful tool to investigate receptor binding of human, alpha-ANP by the antagonizing effect.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D009320:Atrial Natriuretic Factor / Q000032:analysis / Q000276:immunology*; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology*
D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D009320:Atrial Natriuretic Factor