title: Preparation of monoclonal antibodies against atrial natriuretic polypeptide precursor and application to highly sensitive sandwich enzyme immunoassay.
abstract: We produced three monoclonal antibodies against different epitopes of the precursor form of atrial natriuretic polypeptide (ANP), gamma-human ANP (gamma-hANP) [human atrial natriuretic factor (ANF)-(1-126)]. Monoclonal antibodies were prepared by fusion of mouse myeloma cells X63-Ag8.653 with spleen cells from BALB/c mice immunized with synthetic alpha-hANP or gamma-hANP-(1-25) [human ANF-(1-25)] conjugated to thyroglobulin. Of three monoclonal antibodies obtained, two were directed against the alpha-ANP sequence: one antibody (KY-ANP-I) recognizes the N-terminal half of the ring structure, while the other (KY-ANP-II) recognizes the N-terminal sequence of alpha-hANP. The third antibody (KY-ANP-III) was against gamma-hANP-(1-25). With the aid of rabbit polyclonal anti-alpha-ANP-(17-28), we developed sandwich enzyme immunoassays. An enzyme immunoassay for alpha-hANP using KY-ANP-I and polyclonal Fab' was very sensitive (0.01 fmol/tube) and we could detect 0.6 pg/ml of plasma alpha-hANP without extraction. Thus, monoclonal antibodies against gamma-ANP are useful tools for investigating the significance of ANP and related peptides derived from gamma-ANP.
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: K
authors.initials: K
authors.lastname: Hosoda
authors.forename: Y
authors.initials: Y
authors.lastname: Saito
authors.forename: A
authors.initials: A
authors.lastname: Sugawara
authors.forename: H
authors.initials: H
authors.lastname: Arai
authors.forename: G
authors.initials: G
authors.lastname: Shirakami
authors.forename: N
authors.initials: N
authors.lastname: Morii
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D009320:Atrial Natriuretic Factor / Q000276:immunology*; D006801:Humans
chemical_list: D000911:Antibodies, Monoclonal; D009320:Atrial Natriuretic Factor