Investigation of a Radioimmunoassay for Plasma Estrone and Estradiol-17β in Males and Nonpregnant Females. Comparison with an Independent Method using Fluorimetry

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Abstract
A radioimmunoassay for estrone and estradiol-17β in plasma from males and nonpregnant females similar to those used by other workers has been investigated. Chromatography using Sephadex LH-20 separated estrone from estradiol-17β, but neither was separated from other naturally-occurring estrogens. Specifically, estrone was not separated from 2-methoxyestrone and 2-methoxyestradiol-17β, and estradiol-17β was not separated from estradiol-17α, 2-hydroxyestrone, 16-ketoestrone, l6-ketoestradiol-17β, 16α-hydroxyestrone, and 16β-hydroxyestrone. Crossreactions with the antiserum were found for several of these estrogens in both estrone and estradiol-17β assay systems, thus indicating the possibility of substantial interference in the assays. This possibility was investigated. The material measured as plasma estrone and estradiol-17β by radioimmunoasay was subjected to a number of identification procedures and found to have the characteristics of estrone and estradiol-17β. Comparison of plasma estrone and estradiol-17β levels obtained by radioimmunoassay and by a fluorescence procedure which excludes measurement of the other estrogens mentioned above yielded a regression coefficient and intercept not significantly different from 1 and 0, respectively, for both estrone and estradiol-17β determinations. It was concluded that, although the potential for interference remains, other naturally-occurring estrogens do not contribute significantly to the radioimmunoassay values for plasma estrone and estradiol-17β in normal males and nonpregnant females.

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