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Clinical Chemistry 35: 953-957, 1989;
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Clinical Chemistry, Vol 35, 953-957, Copyright © 1989 by American Association for Clinical Chemistry

Development and validation of a two-site immunoradiometric assay for human atrial natriuretic factor in unextracted plasma

HM Lewis, WA Ratcliffe, RA Stott, MR Wilkins and PH Baylis
Department of Pharmacology, University of Birmingham, U.K.

This two-site immunoradiometric assay (IRMA) for human atrial natriuretic factor (ANF)99-126 in plasma utilizes a mouse monoclonal antibody raised against rat (r) ANF103-125 with specificity directed towards the ring structure of ANF, and a rabbit antiserum to human (h) ANF99-126. The monoclonal antibody is radioiodinated, and the IgG fraction of the antiserum is coated onto wells of a microtiter plate. Plasma or standard hANF99-126 (150 microL) is incubated with the radioligand in coated wells for 24 h. The detection limit is 0.9 pg per well, corresponding to 2 pmol/L, with a working range (CV less than 10%) from 4.5 to 540 pmol/L. Intra- and interassay precision are 7% and 9%, respectively, and the assay is unaffected by plasma matrix. In humans, the IRMA is specific for hANF99-126, the major circulating form of ANF, and does not cross-react with metabolites having deletions at the carboxy terminus. Plasma IRMA values in normal seated subjects were 6.6 +/- 1.5 (SEM) pmol/L and results correlated with those of an extraction RIA (r = 0.81, P less than 0.001).


The following articles in journals at HighWire Press have cited this article:


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 Clin. Chem.Home page
A. Clerico, S. Del Ry, and D. Giannessi
Measurement of Cardiac Natriuretic Hormones (Atrial Natriuretic Peptide, Brain Natriuretic Peptide, and Related Peptides) in Clinical Practice: The Need for a New Generation of Immunoassay Methods
Clin. Chem., October 1, 2000; 46(10): 1529 - 1534.
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Copyright © 1989 by the American Association for Clinical Chemistry.