Enzyme immunoassay for 11-ketotestosterone using acetylcholinesterase as laberl: application to the measurement of 11-ketotestosterone in plasma of Siberian sturgeon
Identifieur interne : 001C24 ( Main/Merge ); précédent : 001C23; suivant : 001C25Enzyme immunoassay for 11-ketotestosterone using acetylcholinesterase as laberl: application to the measurement of 11-ketotestosterone in plasma of Siberian sturgeon
Auteurs : B. Cuisset [France] ; P. Pradelles [France] ; D. E. Kime [Royaume-Uni] ; E. R. Kühn [Belgique] ; P. Babin [France] ; S. Davail [France] ; Françoise Le Menn [France]Source :
- Comparative Biochemistry and Physiology, Part C: Comparative Pharmacology and Toxicology [ 0742-8413 ] ; 1994.
Abstract
A competitive enzyme immunoassay using 11-ketotestosterone (11-KT) covalently coupled to acetylcholinesterase (AChE) from electric eel is developed. Ninety-six well microtiter plates are coated with pure mouse-anti-rabbit-IgG monoclonal antibody and competition is performed in one-step reaction by adding successively 11-KT standard or samples, enzymatic tracer and 11-KT-specific antiserum. This assay is more sensitive (detection limit is 7 pg/ml) than radioimmunoassays previously described, and specificity was not modified by the use of enzymatic tracer. This assay can be performed in less than 5 hr. Validation of the EIA was achieved in a fish model, Siberian sturgeon (Acipenser baeri), by comparative determinations of 11-KT levels in plasma extracts using EIA and RIA. Application to the measurement of 11-KT in plasma of farmed Siberian sturgeon during the first reproductive cycle is reported.
Url:
DOI: 10.1016/1367-8280(94)90035-3
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<front><div type="abstract" xml:lang="en">A competitive enzyme immunoassay using 11-ketotestosterone (11-KT) covalently coupled to acetylcholinesterase (AChE) from electric eel is developed. Ninety-six well microtiter plates are coated with pure mouse-anti-rabbit-IgG monoclonal antibody and competition is performed in one-step reaction by adding successively 11-KT standard or samples, enzymatic tracer and 11-KT-specific antiserum. This assay is more sensitive (detection limit is 7 pg/ml) than radioimmunoassays previously described, and specificity was not modified by the use of enzymatic tracer. This assay can be performed in less than 5 hr. Validation of the EIA was achieved in a fish model, Siberian sturgeon (Acipenser baeri), by comparative determinations of 11-KT levels in plasma extracts using EIA and RIA. Application to the measurement of 11-KT in plasma of farmed Siberian sturgeon during the first reproductive cycle is reported.</div>
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