Hapten Synthesis and Antibody Development for Polychlorinated Dibenzo-p-dioxin Immunoassays

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• 作者：James R. Sanborn ; Shirley J. Gee ; S. Douglass Gilman ; Yukio Sugawara ; A. Daniel Jones ; Jane Rogers ; Ferenc Szurdoki ; Larry H. Stanker ; Donald W. Stoutamire ; and Bruce D. Hammock
• 刊名：Journal of Agricultural and Food Chemistry
• 出版年：1998
• 出版时间：June 1998
• 年：1998
• 卷：46
• 期：6
• 页码：2407 - 2416
• 全文大小：161K
• 年卷期：v.46,no.6(June 1998)
• ISSN：1520-5118

文摘

This paper reports the synthesis of haptens and the generation andpreliminary evaluation ofpolyclonal antibodies for the detection of dioxins such as TCDD(2,3,7,8-tetrachlorodibenzo-p-dioxin)by ELISA (enzyme-linked immunosorbent assay). These novel haptenscontain unsaturation betweenthe halogenated dibenzo-p-dioxin ring system and the proteinto which it is conjugated, presentinga rigid handle structure. The substitution pattern is identicalwith or similar to that of TCDD (i.e.,2,3,7,8- or 1,2,3,7,8-). Finally, the haptens lack polar groupsfor hydrogen bonding. In direct bindingassays using the new polyclonal antibodies there was excellentrecognition of hapten-proteinconjugates, including recognition of those hapten conjugates that werenot used as immunogens(i.e., assay systems heterologous in hapten structure). Thesehaptens do elicit selective immuneresponses in rabbits. Their evaluation in an ELISA formatdemonstrated the usefulness of thesehaptens for the detection of dioxins. An IC₅₀ of 0.8ng/well (16 ng/mL) was observed for anunoptimized system that used2,3,7-trichloro-8-methyldibenzo-p-dioxin as an analyticalsurrogatestandard.Keywords: TCDD; dioxin; immunoassay; polyclonal antibodies; haptensynthesis; polychlorinatedhydrocarbons