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Chemiluminescence immunoassays for estradiol and ethinylestradiol based on new biotinylated estrogen derivatives

Kanso, Hussein, Inguimbert, Nicolas, Istamboulie, Georges, Barthelmebs, Lise, Calas-Blanchard, Carole, Noguer, Thierry
Analytical biochemistry 2017 v.537 pp. 63-68
antibodies, biotin, chemical bonding, chemiluminescence immunoassays, effluents, estradiol, hydroxyl radicals, peroxidase, serum albumin, wastewater treatment
New chemiluminescence-based immunoassays for sensitive detection of 17-β estradiol (E2) and ethinylestradiol (EE2) are described on the basis of the use of biotinylated estrogen derivatives. Estrogen derivatives bearing a carboxylic group (E2-COOH and EE2-COOH) on C-3 position were synthesized, covalently bound to aminated biotin and subsequently immobilized on avidin-coated microtiter plates. The assay principle was based on competition between free and immobilized estrogens for their binding to primary antibodies, with subsequent revelation using horseradish peroxidase (HRP)-labeled secondary antibodies. Under optimized conditions, the chemiluminescence immunoassays showed a highly sensitive response to E2 and EE2, with respective detection limits of 0.5 and 1.2 ng L−1. The LOD achieved using biotinylated E2 was in the same order of magnitude as those obtained using commercially available E2-bovine serum albumin conjugate (E2-BSA). The developed devices were successfully applied to analysis wastewater treatment plants effluents (WWTP) with negligible matrix effect.