Radioimmunoassay of estradiol-17$beta$ without chromatography

Journal Article · Tue Jan 01 04:00:00 EST 1974 · J. Clin. Endocrinol. Metab., v. 38, no. 1, pp. 42-50

DOI:https://doi.org/10.1210/jcem-38-1-42· OSTI ID:4365374

England, B G; Niswender, G D; Midgley, Jr, A R

Several radioimmunoassays for the estimation of estradiol-17 BETA in serum have been reported. For the generation of antisera, the majority of these methods have used steroids conjugated to a carrier protein through sites on the A or D rings. The resultlng antisera have shown considerable cross reactivity with related steroids necessitating the use of various chromatographic separation steps prior to the actual radioimmunoassay. The use of haptens in which the carrier protein was conjugated to a steroid molecule through carbon 6 on the B ring (1,2,3) has resulted in the production of highly specific antisera. Conjugation of the protein molecule to other positions which leave the A and D rings available for recognition by antibody should also result in the generation of antibodies with enhanced specificity. In this report, an antiserum was prepared to a conjugate of an 11 BETA -hemisuccinate derivative of estradiol-17 BETA to bovine serum albumin. The resulting antiserum appeared to be specific for 17 BETA -estradiol and was sensitive to substitutions on the A, B and D rings. With the use of this antiserum, it has been possible to develop a sensitive, accurate and specific radioimmunoassay for the estimation of estradiol-17 BETA in sera from humans. This artiserum was sufficiently specific to allow the use of a simple benzene extract of serum without subsequent chromatography. The use of iodinated E/sub 2/- BETA -11 alpha -TME as labeled tracer further simplified the procedure by allowing the use of gamma ray spectrometry, rather than liquid scintillation counting with its attendant expense and inconvenience. (auth)

• Research Organization: Univ. of Michigan, Ann Arbor
• Sponsoring Organization: USDOE
• NSA Number: NSA-29-018747
• OSTI ID: 4365374
• Journal Information: J. Clin. Endocrinol. Metab., v. 38, no. 1, pp. 42-50, Journal Name: J. Clin. Endocrinol. Metab., v. 38, no. 1, pp. 42-50; ISSN JCEMA
• Country of Publication: Country unknown/Code not available
• Language: English

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