Parallel radioimmunoassay for plasma cortisol and 11-deoxycortisol

Abstract

The authors describe a direct, rapid, and specific procedure for the parallel radioimmunoassay for cortisol and 11 deoxycortisol in plasma. The plasma sample is used directly, after heat inactivation of the natural cortisol binding protein. The radioimmunoassay utilizes antibodies generated in rabbits by steroids conjugated at their 3 oxo position to thyroglobulin. Ammonium sulfate is used to separate bound and free steroids. Our cortisol antibody and an 11 deoxycortisol antibody obtained elsewhere cross reacted negligibly with each other or with other steroids that might be present in plasma. Radioimmunoassays were therefore developed for both steroids in only 1.25 μl of plasma. The intra and inter assay coefficients of variation for both steroids were less than 10%, with a sensitivity of 4 μg/liter. Steroid values obtained by a competitive protein binding method were consistently higher than those of the present method, suggesting that the former is measuring total corticosteroids. This simple approach requires only 4 h for the specific measurement of both cortisol and 11 deoxycortisol in 20 samples of plasma.