A monoclonal antibody against N-terminus of α-atrial natriuretic polypeptide (α-ANP): A useful tool for preferential detection of naturally circulating ANP

M. Mukoyama, K. Nakao, T. Yamada, H. Itoh, A. Sugawara, Y. Saito, H. Arai, K. Hosoda, G. Shirakami, N. Morii, S. Shiono, H. Imura

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Abstract

A monoclonal antibody directed against the N-terminus of α-atrial natriuretic polypeptide (α-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 α-human ANP (α-hANP) conjugated to bovine thyroglobulin. The obtained antibody showed a high affinity for α-hANP with a Ka of 6.6 x 1010 M-1. With this monoclonal antibody, a specific radioimmunoassay for α-hANP was established. The minimal detectable level of α-hANP in this radioimmunoassay was 0.8 fmol (2.5 pg)/tube, and IC50 was 8 fmol (25 pg)/tube. The radioimmunoassay recognized α-rat ANP on an equimolar basis, whereas there was no detectable cross-reactivity with α-ANP [4-28] or α-ANP [5-28]. Thus, the monoclonal antibody can detect a hormonal form of ANP, α-ANP, but not neuropeptide forms of ANP, α-ANP [4-28] and α-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.

Original languageEnglish
Pages (from-to)1277-1284
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume151
Issue number3
Publication statusPublished - 1988 Jan 1

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Mukoyama, M., Nakao, K., Yamada, T., Itoh, H., Sugawara, A., Saito, Y., Arai, H., Hosoda, K., Shirakami, G., Morii, N., Shiono, S., & Imura, H. (1988). A monoclonal antibody against N-terminus of α-atrial natriuretic polypeptide (α-ANP): A useful tool for preferential detection of naturally circulating ANP. Biochemical and Biophysical Research Communications, 151(3), 1277-1284.