A Simplified Radioimmunoassay of Human Gravid Plasma Estriol

概要

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A highly simplified and specific radioimmunoassay (RIA) method was developed to determine plasma unconjugated estriol (E3) during pregnancy. The antigen used for immunization of rabbits was E3-6-0-carboxymethyp-oxime-bovine serum albumin (E3-6-oxime-BSA), a derivative of 6-oxo-E3. At 24 weeks after the first immunization, the antiserum diluted 1: 50, 000 showed almost no cross reaction with neutral steroids and little cross reaction with phenolic steroids except for estrone-3-sulfate (32.6%). Plasma samples were extracted with ethyl ether and evaporated to dryness. No chromatographic purification was necessary. The dried extracts were incubated with antiserum at 24°C for 30min, then antibody-bound and free E3 were separated with dextran coated charcoal. The sensitivity of this method was 10pg. The intra and inter coefficients of variation were 7.8% and 11.6%. The coefficient of variation of the present method was 12.8% compared to a method using a LH-20 column chromatography for purification. Using our simplified method, the mean plasma E3 during pregnancy was 2.6±1.4ng/ml at 16-20 weeks and 16.4±5.0ng/ml at term.
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