A NEW RADIOIMMUNOASSAY FOR HUMAN ALPHA-ATRIAL NATRIURETIC PEPTIDE AND ITS PHYSIOLOGICAL VALIDATION

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 x 10111/mol) showed no cross-reactivity with related analogues. Displacement of 50% bound 12^I-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). © 1990, Marcel Dekker, Inc.