A New Radioimmunoassay for Human Alpha Atrial Natriuretic Peptide and Its Physiological Validation

Abstract

A new sensitive, specific radioimmunoassay for human alpha atrial natriuretic peptide (hANP) and a novel extraction method for hANP have been developed. Antiserum to hANP (Keq = 1.923 × 10¹¹ 1/mol) showed no cross-reactivity with related analogues. Displacement of 50% bound ¹²⁵I-hANP occurred at 4.7 ± 0.1 fmol/tube (n=15). The limit of detection of plasma hANP after extraction of 2ml plasma with Florisil was 1.2 pmol hANP/litre plasma. The recovery of synthetic hANP from plasma over the range 6.5–162.5 pmol/1 was 71.2 ± 1.9%. Inter- and intra-assay coefficients of variation were 13.1% and 10.1% respectively. Extracted plasma was stored at −80°C without loss in immunoreactivity. The radioimmunoassay was physiologically validated in man by measuring plasma hANP following central volume expansion -

{a} plasma hANP rose from 2.5 ± 0.5 to 4.1 ± 0.6 pmol/1 in 9 normal volunteers after postural change from seated to lying

{b} infusion of normal saline (2 litre/2 hour) increased hANP from 1.6 to 4.3 pmol/1 (n=2).

Infusion of hANP (2pmol/kg/min) increased plasma hANP to 19.9 ± 3.4 pmol/1 (n=6).