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Development of Magnetic Particle-based Chemiluminescence Enzyme Immunoassay for the Detection of 17β-estradiol in Environmental Water

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Abstract

In the present work, a simple, fast, and highly sensitive chemiluminescence enzyme immunoassay for 17β-estradiol (E2) in environmental water samples was developed, using magnetic particles (MPs) labeled with secondary antibody as both the immobilization matrix and the separation tools. The specific anti-E2 polyclonal antibody (PcAb) was produced against a conjugate of estradiol–bovine serum albumin. The specificity of the anti-E2 antibody was studied. The results showed that the antibody did not cross-react with the structurally related endocrine-disrupting compounds, including estrone, ethinyl E2, estriol, E2-17-glucuronide, E2-3-sulfate-17-glucuronide, androstenedione, and dihydrotestosterone. The water samples were pretreated with solid-phase extraction using C18 cartridges for the removal of matrix effects. Several physicochemical parameters including the dilution ratios of E2-6–horseradish peroxidase conjugate and anti-E2 PcAb, immunoreaction time, volume of chemiluminescent substrate and MPs, chemiluminescence reaction time, and pH of assay solution were studied and optimized. At optimal experimental conditions, it was found that the proposed method exhibited high performance with detection limit of 2.0 pg/mL, linear range of 20–1,200 pg/mL, and total assay time of 45 min. Both inter- and intra-assay coefficient of variation were less than 10%. The average recoveries of three different spiked concentration samples ranged from 86.3% to 108%. The method was successfully applied to the determination of E2 in river, waste, and tap water, and showed a good correlation with the commercially available radioimmunoassay kit.

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Acknowledgements

This work was supported by the National Basic Research Program of China (973 Program, No. 2007CB714507) and National Nature Science Foundation of China (No. 20728505).

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Authors and Affiliations

  1. The Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Department of Chemistry, Tsinghua University, Beijing, 100084, China

    Tian-Bing Xin, Xu Wang, Hui Jin & Jin-Ming Lin

  2. College of Chemistry and Environmental Science, Hebei University, Baoding, 071002, China

    Tian-Bing Xin, Hui Jin & Shu-Xuan Liang

  3. Beijing Chemclin Biotech Co. Ltd, Beijing, 100094, China

    Zhen-Jia Li

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  1. Tian-Bing Xin
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Correspondence to Xu Wang or Jin-Ming Lin.

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Xin, TB., Wang, X., Jin, H. et al. Development of Magnetic Particle-based Chemiluminescence Enzyme Immunoassay for the Detection of 17β-estradiol in Environmental Water. Appl Biochem Biotechnol 158, 582–594 (2009). https://doi.org/10.1007/s12010-008-8356-3

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