Preparation of monoclonal antibodies against atrial natriuretic polypeptide precursor and application to highly sensitive sandwich enzyme immunoassay

Masashi Mukoyama, Kazuwa Nakao, Takayuki Yamada, Hiroshi Itoh, Kiminori Hosoda, Yoshihiko Saito, Akira Sugawara, Hiroshi Arai, Gotaro Shirakami, Narito Morii, Shozo Shiono, Seiichi Hashida, Eiji Ishikawa, Hiroo Imura

研究成果: Article査読

3 被引用数 (Scopus)

抄録

We produced three monoclonal antibodies against different epitopes of the precursor form of atrial natriuretic polypeptide (ANP), γ -human ANP (γ-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 α-hANP or γ -hANP-(1-25) [human ANF-(1-25)] conjugated to thyroglob- ulin. Of three monoclonal antibodies obtained, two were directed against the α -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 α -hANP. The third antibody (KY-ANP-III) was against γ -hANP-(1-25). With the aid of rabbit polyclonal anti-α-ANP-(17-28), we developed sandwich enzyme immunoassays. An enzyme immunoassay for a-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 α -hANP without extraction. Thus, monoclonal antibodies against γ -ANP are useful tools for investigating the significance of ANP and related peptides derived from γ-ANP.

本文言語English
ページ(範囲)S320-S322
ジャーナルJournal of Hypertension, Supplement
6
4
DOI
出版ステータスPublished - 1988 12
外部発表はい

ASJC Scopus subject areas

  • 内科学
  • 生理学
  • 循環器および心血管医学

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