Development and evaluation of a simple, direct, solid-phase radioimmunoassay of serum cortisol from readily available reagents
A simple, rapid solid-phase radioimmunoassay for serum cortisol was developed using cortisol antibody distributed by the Scottish Antibody Production Unit and commercially available radioiodinated cortisol ligand. The assay involves a 1-h incubation at ambient temperature, using the antibody covalen...
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Veröffentlicht in: | Clinica chimica acta 1986-07, Vol.158 (1), p.59-70 |
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Sprache: | eng |
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Zusammenfassung: | A simple, rapid solid-phase radioimmunoassay for serum cortisol was developed using cortisol antibody distributed by the Scottish Antibody Production Unit and commercially available radioiodinated cortisol ligand. The assay involves a 1-h incubation at ambient temperature, using the antibody covalently linked by the easily performed carbonyldiimidazole method, to microcrystalline cellulose.
A detailed comparison of the accepted 0.125 mol/l citrate, pH 4.0, and an alternative 0.1 mol/l phosphate/8-anilinonaphthalene sulphonic acid, pH 7.4, diluent demonstrated similar precision (within-assay 10% to 5%; between-assay 11% to 6% over the range 100–1000 nmol/l cortisol) and recovery (median recoveries: phosphate 98.6%, citrate 99.5%). Linear regression analysis of human serum cortisol concentrations in comparison with the ‘Amerlex’ cortisol RIA kit gave the following results: citrate = 1.029. ‘Amerlex’ - 3.5 nmol/l (
n = 80,
r = 0.967); phosphate = 1.017, ‘Amerlex’+ 26.2 nmol/l (
n = 80,
r = 0.961).
Phosphate, pH 7.4 diluent was adopted as the diluent of choice, since it was economical of antibody and maintained good precision over a wider working range of cortisol concentration. |
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ISSN: | 0009-8981 1873-3492 |
DOI: | 10.1016/0009-8981(86)90116-6 |