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Journal of Clinical Endocrinology & Metabolism Vol. 44, No. 1 91-95
doi:10.1210/jcem-44-1-91
Copyright © 1977 by the Endocrine Society.
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Enzyme-Labelled Immunoassay for Plasma Cortisol

TOSHIO OGIHARA, KIYOSHI MIYAI*, KEIKO NISHI*, KAICHIRO ISHIBASHI** and YUICHI KUMAHARA

* Department of Medicine and Geriatrics, the Central Laboratory for Clinical Investigation Tokyo, Japan
** Osaka University Medical School, Osaka and Eiken Immunochemical Institute Tokyo, Japan

Correspondence should be addressed to: Kiyoshi Miyai, M.D., The Central Laboratory for Clinical Investigation,Osaka University Hospital, Fukushima-ku,Osaka 553, Japan.

An enzyme-labelled immunoassay for plasma cortisol was developed. For this alkalinephosphatase was conjugated through 21-hemisuccinate of cortisol using water-soluble carbodiimide. An antibody was raised in rabbits against cortdsol-21-hemisuccinate bovine serum albumin. Before assay 10 βl samples of plasma were mixed with glutamate buffer, pH 3.3, and boiled to denature endogenous cortisol-binding globulin and alkalinephosphatase. Separation of "bound and free" cortisol was done by the double antibody method. A linear relationship was obtained between the volume of plasma and the amount of cortisol. The minimal detectable level of plasma cortisol was 1 µg/dl and the coefficients of variation were 2.7%–4.4% (within assays) and 4.7%–6.0% (between assays). Cortisol values determined by the present method correlated well with those determined by radioimmunoassay. The present method of enzyme-labelled immunoassay is suitable for routine clinical analysis of plasma cortisol.

Supported by grants from the Ministry of Education and the Ministry of Health and Welfare, Japan.

Received April 26, 1976.






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Copyright © 1977 by The Endocrine Society