Development of a highly sensitive, second antibody format chemiluminescence enzyme immunoassay for the determination of 17β-estradiol in wastewater

In the present work, a highly sensitive chemiluminescence enzyme immunoassay (CLEIA) method for the determination of 17 beta-estradiol (E2) in the wastewater was developed, which used the fluorescein-iso-thiocyanate (FITC)-anti-FITC system as solid phase and the biotin-streptavidin system as signal amplification system, with the highly sensitive 4-methoxy-4-(3-phosphatephenyl)-spiro-(1,2-dioxetane-3,2'-adamantane) (AMPPD)-alkaline phosphatase (ALP) system as chemiluminescence detection system. The specific polyclonal antibody was produced against a conjugate of estradiol-bovine serum albumin (BSA), biotinoyl-diaminodioxaoctane-estradiol conjugate was synthesized. The specificity of anti-E2 antibody was also studied. The results showed that anti-E2 antibody has no cross-reactions for estriol, ethinyl E2, estrone commonly coexisting with E2 in the wastewater. Samples were prepared with solid-phase extraction using C-18 cartridges for the removal of matrix effects. The linear range was 2.5-1600 pg ml(-1) and detection limit was 1.5 pg ml(-1). As our known of knowledge, the method was more sensitive for the determination of E2 in water compared with other immunoassay methods. Recoveries were in the range of 80-110% in spiked tap water and wastewater. This method has been successfully used for the detection of E2 in wastewater, compared with the commercially radioimmunoassay (RIA) kit, the correlation was good. (c) 2005 Elsevier B.V. All rights reserved.