Development and Utilization of Camelid VHH Antibodies from Alpaca for 2,2 ',4,4 '-Tetrabrominated Diphenyl Ether Detection

文献类型： 外文期刊

第一作者： Bever, Candace R. S.

作者： Bever, Candace R. S.;Majkova, Zuzana;McCoy, Mark;Wang, Yanru;Gee, Shirley;Hammock, Bruce D.;Radhakrishnan, Rajeswaran;Suni, Ian;McCoy, Mark;Wang, Yanru;Dechant, Julie;Hammock, Bruce D.

作者机构：

期刊名称：ANALYTICAL CHEMISTRY （ 影响因子：6.986； 五年影响因子：6.755 ）

ISSN： 0003-2700

年卷期： 2014 年 86 卷 15 期

页码：

收录情况： SCI

摘要： An antibody-based analytical method for the detection of a chemical flame retardant using antibody fragments isolated from an alpaca has been developed. One specific chemical flame retardant congener, 2,2',4,4'-tetrabrominated diphenyl ether (BDE-47), is often the major poly-BDE (PBDE) congener present in human and environmental samples and that which is the most frequently detected. An alpaca was immunized with a surrogate of BDE-47 covalently attached to a carrier protein. The resulting mRNA coding for the variable domain of heavy-chain antibodies (VHH) were isolated, transcribed to cDNA, and cloned into a phagemid vector for phage display library construction. Selection of VHHs recognizing BDE-47 was achieved by panning under carefully modified conditions. The assay sensitivity for detecting BDE-47 was down to the part-per-billion (microgram per liter) level. Cross-reactivity analyses confirmed that this method was highly selective for BDE-47 and selected hydroxylated metabolites. When exposed to elevated temperatures, the camelid VHH antibodies retained more reactivity than a polyclonal antibody developed to the same target analyte. The use of this VHH antibody reagent immobilized onto a Au electrode for impedance biosensing demonstrates the increased versatility of VHH antibodies.

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