Radioimmunoassay of 17-Hydroxyprogesterone in Serum with or without Thin-Layer Chromatographic Purification

Abstract

A radioimmunoassay for the measurement of 17-hydroxyprogesterone (17-OHP) is described. The antigen ll-desoxycortisol-21-hemisuccinate-bovine serum albumin has been used to produce two antisera of different antibody populations in the same animal. The thin-layer chromatographic system described can be used to separate all the cross-reacting steroids investigated from 17-OHP. A simplified method is also presented using preliminary solvent extraction only. The mean 17-OHP levels measured, using this method, were, for normal men, 0.86 ± 0.19 ng/ml (SD), for normal females 0.30 ± 0.19 ng/ml in the follicular phase and 1.72 ± 0.18 ng/ml in the luteal phase of the menstrual cycle, and 0.45 ± 0.17 ng/ml in normal postmenopausal women.

Use of Highly Specific Antibodies against 17α-OH-Progesterone in a Simplified Nonchromatographic RIA and in the Simultaneous Determination of Four Sex Hormones in Human Plasma

“…Our results are in good agreement with those previously reported using radio-ligands with markedly different specific requirements (1,6,16,18,20,23) or RIA with different chromatographic purifications (2,10,22,24).…”

“…19),hydroxyalkoxypropyl Sephadex (LH-20) (6) or a single purification step by paper chro matography (18), liquid gel chromatography on Sephadex 15,22,24) Forest 261 and its hydrophobic derivative (8,10,11). Methods using unchromatographed serum extracts without any further purification (7) or with an 'immunological' purification (23) prior to radioimmunological measurements utilizing relatively unspecific antisera have also been proposed.…”

A Simple Procedure for the Radioimmunoassay of 17α-Hydroxyprogesterone in Serum: Comparison with an Immunological Purification Technique

“…In contrast to other reports (Abraham et al, 1971;Wickings, 1975), the mean concentration of 17-0HP in the serum of women in the luteal phase of the menstrual cycle was significantly higher than that during the follicular phase. Youssefnejadian et al (1972), as in the present study, were able to distinguish between a lower mean level of 17-0HP (days 4-8 of the cycle) and the mean level found for days 18-28 of the cycle in pools of serum from healthy women.…”

“…Further studies with the simple extraction method are required to assess whether the technique will distinguish the midcycle peak of 17-0HP as demonstrated by Hami et al (1975) and Aedo et al (1976). Hami et al (1975) Wickings (1975 …”

“…A number of radioimmunoassay methods have been described (Abraham et al, 1971;Youssefnejadian et al, 1972;Franks, 1974;Hami et al, 1975;Janne et al, 1975;Wickings, 1975;Kaplan, 1977;Forest and Cathiard, 1978;Hughes and Winter, 1978;Parth et al, 1978), but in the majority various degrees of purification of the serum extract are considered necessary. This detracts from the value of these methods when a result is required urgently.…”