title: A monoclonal antibody against N-terminus of alpha-atrial natriuretic polypeptide (alpha-ANP): a useful tool for preferential detection of naturally circulating ANP.
abstract: A monoclonal antibody directed against the N-terminus of alpha-atrial natriuretic polypeptide (alpha-ANP), named KY-ANP-II, was produced by fusion of a non-producing mouse myeloma cell line, X63-Ag8.653, with spleen cells from a BALB/c mouse immunized with synthetic alpha-human ANP (alpha-hANP) conjugated to bovine thyroglobulin. The obtained antibody showed a high affinity for alpha-hANP with a Ka of 6.6 X 10(10) M-1. With this monoclonal antibody, a specific radioimmunoassay for alpha-hANP was established. The minimal detectable level of alpha-hANP in this radioimmunoassay was 0.8 fmol (2.5 pg)/tube, and IC50 was 8 fmol (25 pg)/tube. The radioimmunoassay recognized alpha-rat ANP on an equimolar basis, whereas there was no detectable cross-reactivity with alpha-ANP [4-28] or alpha-ANP [5-28]. Thus, the monoclonal antibody can detect a hormonal form of ANP, alpha-ANP, but not neuropeptide forms of ANP, alpha-ANP [4-28] and alpha-ANP [5-28]. These results indicate that KY-ANP-II becomes a useful tool for preferential detection of a circulating form of ANP and for investigation of the physiological and pathophysiological significance of ANP as a hormone.
authors.affiliation: Department of Medicine, Kyoto University School of Medicine, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Mukoyama
authors.forename: K
authors.initials: K
authors.lastname: Nakao
authors.forename: T
authors.initials: T
authors.lastname: Yamada
authors.forename: H
authors.initials: H
authors.lastname: Itoh
authors.forename: A
authors.initials: A
authors.lastname: Sugawara
authors.forename: Y
authors.initials: Y
authors.lastname: Saito
authors.forename: H
authors.initials: H
authors.lastname: Arai
authors.forename: K
authors.initials: K
authors.lastname: Hosoda
authors.forename: G
authors.initials: G
authors.lastname: Shirakami
authors.forename: N
authors.initials: N
authors.lastname: Morii
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D009320:Atrial Natriuretic Factor / Q000097:blood*; D003429:Cross Reactions; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology*; D011863:Radioimmunoassay
chemical_list: D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; D009320:Atrial Natriuretic Factor