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  • Title: Preparation of monoclonal antibodies against atrial natriuretic polypeptide precursor and application to highly sensitive sandwich enzyme immunoassay.
    Author: Mukoyama M, Nakao K, Yamada T, Itoh H, Hosoda K, Saito Y, Sugawara A, Arai H, Shirakami G, Morii N.
    Journal: J Hypertens Suppl; 1988 Dec; 6(4):S320-2. PubMed ID: 2977170.
    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.
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