Development of a highly sensitive enzyme-linked immunosorbent assay based on polyclonal antibodies for the detection of polychlorinated dibenzo-p-dioxins

Analytical Chemistry
Y SugawaraB D Hammock

Abstract

The development of an enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibodies for the polychlorinated dibenzo-p-dioxins is described. We previously reported the synthesis of haptens and generation of antibodies for detection of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Antisera were screened with seven different coating antigens (hapten-protein conjugates), including trans-3-(7,8-dichlorodibenzo-p-dioxin-2-yl)-cis-2-methylpropeno ic acid (VII) and 5-(3,7,8-trichlorodibenzo-p-dioxin-2-yl)penta-trans,trans-2,4-dien oic acid (X). All inhibition screening and optimization studies were conducted using a less toxic surrogate standard for TCDD [2,3,7-trichloro-8-methyl-dibenzo-p-dioxin (TMDD; XVII)] which responded similarly to 2,3,7,8-TCDD in the ELISA. The most sensitive assay from the screening studies [coating antigen VII-BSA, 0.1 microgram/mL, and antiserum 7598 (anti-X-LPH), 1:10,000] was further optimized and characterized. It exhibited an IC50 value of 12 pg/well (240 pg/mL), with working range from 2 to 240 pg/well (40 to 4800 pg/mL). The influence of various physical and chemical factors (time, solvent, detergent) was investigated. The optimized assay was then used to assess cross-reactivity by congeners o...Continue Reading

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Citations

Jan 22, 2010·Archives of Environmental Contamination and Toxicology·E García-NietoF Díaz-Barriga
Jun 7, 2002·The Science of the Total Environment·Mark E Hahn
Nov 26, 1999·The Science of the Total Environment·R O Harrison, G H Eduljee
Mar 8, 2000·Proceedings of the National Academy of Sciences of the United States of America·G ShanB D Hammock
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Mar 5, 2004·Biopolymers·Chikashi NakamuraJun Miyake
Apr 19, 2011·Journal of Environmental Science and Health. Part. B, Pesticides, Food Contaminants, and Agricultural Wastes·Yi H LiuGuo N Zhu
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Dec 20, 1999·Journal of Agricultural and Food Chemistry·G ShanB D Hammock

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