血漿コルチゾールのエンザイムイムノアッセイ-基礎的検討と臨床応用

概要

論文の詳細を見る
We have developed a clinically useful enzyme-labelled immunoassay (EIA) for plasma cortisol. Alkaline-phosphatase (AL-P) from E. coli was used as a tracer. Water soluble carbodiimide was used as a conjugating reagent between enzyme and cortisol-21-hemisuccinate. Cortisol-AL-P conjugate was separated from unreacted cortisol by means of a Sephadex G-100 column. Anti-cortisol-21-hemisuccinate BSA rabbit serum was used in this system, however anti-cortisol-3-oxime BSA rabbit serum did not work in this system. 10 μl of plasma was used for assay after denaturation of endogenous cortisol binding globulin and AL-P by heating in a glutamate buffer. B-F separation was done by means of the double antibody method. AL-P activity of antigen-antibody complex was measured by means of the Kind-King method. Sensitivity, specificity and measurable range of the present EIA method were quite similar to those of the RIA method using 125I-cortisol or 3H-cortisol. This method might be useful and generally available for routine analysis of plasma cortisol because the use of radioactive materials is avoided.
日本内分泌学会の論文