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

Journal Article · Thu Jun 01 00:00:00 EDT 1989 · Clin. Chem. (Winston-Salem, N.C.); (United States)

OSTI ID:5934326

Lewis, H M; Ratcliffe, W A; Stott, R A; Wilkins, M R; Baylis, P H

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).

• Research Organization: Univ. of Birmingham (England)
• OSTI ID: 5934326
• Journal Information: Clin. Chem. (Winston-Salem, N.C.); (United States), Vol. 35:6
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PEPTIDES
RADIOIMMUNOASSAY
IMMUNOGLOBULINS
IODINE ISOTOPES
MAN
MICE
MONOCLONAL ANTIBODIES
RABBITS
SPECIFICITY
ANIMALS
ANTIBODIES
BIOASSAY
GLOBULINS
IMMUNOASSAY
IMMUNOLOGY
ISOTOPE APPLICATIONS
ISOTOPES
MAMMALS
ORGANIC COMPOUNDS
PRIMATES
PROTEINS
RADIOASSAY
RADIOIMMUNOLOGY
RODENTS
TRACER TECHNIQUES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques