A simple, direct radioimmunoassay for plasma cortisol, featuring a 125I radioligand and a solid-phase separation technique

Abstract

An antiserum, raised in sheep to a cortisol-3-(O-carboxymethyl)oxime/bovine serum albumin conjugate, is coupled to microcellulose. No extraction is required because plasma samples and standards are incubated with the antiserum and an 125I radioligand in a low-pH buffer, which denatures cortisol-binding globulins. The assay satisfies accepted validation criteria. In addition, results from the radioimmunoassay compare well with those obtained by a gas chromatographic-mass spectrometric technique (r = 0.968; F(RIA) = 0.97 F(GCMS) + 2.0 nmol/L). The latter procedure features the very high intrinsic specificity obtained by selected ion monitoring at high mass-spectrometric resolution (M/ΔM = 8500) with a Varian MAT-731 instrument. The simplicity of the radioimmunoassay procedure, with use of reagents prepared 'in house', makes this a very practical and economical assay for use in the medium or large endocrine laboratory.