Preparation of conjugates of progesterone with bovine serum albumin in the reversed micellar medium

Yatsimirskaya, Eugenia A.; Gavrilova, E. M.; Egorov, A. M.; Levashov, Andrey V.

doi:10.1016/0039-128x(93)90033-j

Cited by 41 publications

(14 citation statements)

References 10 publications

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“…The immunogen was synthesized according to Yatsimirskaya et al with minor modifications (31). In brief, 12 mg of the 5Ј-O-carboxymethoxy Enl, was left to react overnight with DCC and NHS (molar ratio, 3:4:5) in 200 l of anhydrous dimethylformamide.…”

Section: Fig 1 Structures Of Mammalian Lignans and Their Precursorsmentioning

confidence: 99%

Time-Resolved Fluoroimmunoassay for Plasma Enterolactone

Adlercreutz

Wang

Lapčı́k

et al. 1998

Analytical Biochemistry

135

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Section: Fig 1 Structures Of Mammalian Lignans and Their Precursorsmentioning

confidence: 99%

Time-Resolved Fluoroimmunoassay for Plasma Enterolactone

Adlercreutz

Wang

Lapčı́k

et al. 1998

Analytical Biochemistry

135

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“…The absorbances of the conjugates were compared with that of HSA at the same concentrations, and calculations were performed by the method of Erlanger et al [36] using a molecular weight of HSA as 66,462 g mol −1 and the extinction coefficient (ε) for 17, which was assumed to remain the same when the steroids were conjugated to HSA. While this assumption is reported to give a relative error of ∼10% due to the shift in the steroid 17 absorption maximum upon conjugation [38,39], it remains constant for conjugates of a given type, thus enabling comparisons of the same conjugates produced by different methods [40]. The results of these steroid-HSA molar ratio calculations are shown in Table 1 for 23a, 23b and 26a.…”

Section: Synthesismentioning

confidence: 97%

Analysis of anabolic steroids in the horse: Development of a generic ELISA for the screening of 17α-alkyl anabolic steroid metabolites

Hungerford

Sortais

Smart

et al. 2005

The Journal of Steroid Biochemistry and Molecular Biology

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Due to the potential for misuse of a wide range of anabolic steroids in horse racing, a screening test to detect multiple compounds, via a common class of metabolites, would be a valuable forensic tool. An enzyme-linked immunosorbent assay (ELISA) has been developed to detect 17alpha-alkyl anabolic steroid metabolites in equine urine. 16beta-Hydroxymestanolone (16beta,17beta-dihydroxy-17alpha-methyl-5alpha-androstan-3-one) was synthesised in six steps from commercially available epiandrosterone (3beta-hydroxy-5alpha-androstan-17-one). Polyclonal antibodies were raised in sheep, employing mestanolone (17beta-hydroxy-17alpha-methyl-5alpha-androstan-3-one) or 16beta-hydroxymestanolone conjugated to human serum albumin, via a 3-carboxymethyloxime linker, as antigens. Antibody cross-reactivities were determined by assessing the ability of a library of 54 representative steroids to competitively bind the antibodies. Antibodies raised against 16beta-hydroxymestanolone showed excellent cross-reactivities for all of the 16beta,17beta-dihydroxy-17alpha-methyl steroids analysed and an ELISA has been developed to detect these steroid metabolites. Using this 16beta-hydroxymestanolone assay, urine samples from horses administered with stanozolol (17alpha-methyl-pyrazolo[4',3':2,3]-5alpha-androstan-17beta-ol), were analysed raw, following beta-glucuronidase hydrolysis, and following solid-phase extraction (SPE) procedures. The suppressed absorbances observed were consistent with detection of the metabolite 16beta-hydroxystanozolol. Positive screening results were confirmed by comparison with standard LCMS analyses. Antibodies raised against mestanolone were also used to develop an ELISA and this was used to detect metabolites retaining the parent D-ring structure following methandriol (17alpha-methylandrost-5-ene-3beta,17beta-diol) administration. The ELISA methods developed have application as primary screening tools for detection of new and known anabolic steroid metabolites.

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“…The common mixed anhydride, and the micellar method of Yatsimirskaya et al [10] have been used for conjugation of each hapten to bovine serum albumin (BSA). The starting hapten-protein molar ratio was 50, the resulting conjugates contained 10-12 and 11-14 mol of the hapten in 1 mol of the conjugates, respectively, as determined by spectrophotometry with trinitrobenzene sulfonic acid.…”

Section: Preparation Of Immunogensmentioning

confidence: 99%