Development and utilization of camelid VHH antibodies from alpaca for 2,2',4,4'-tetrabrominated diphenyl ether detection.
Development and utilization of camelid VHH antibodies from alpaca for 2,2',4,4'-tetrabrominated diphenyl ether detection.
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DOI：
10.1021/ac501807j
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发表时间：
2014-08-05
影响因子：
7.4
通讯作者：
Hammock, Bruce D.
中科院分区：
文献类型：
--
作者：
Bever, Candace R. S.;Majkova, Zuzana;Radhakrishnan, Rajeswaran;Suni, Ian;McCoy, Mark;Wang, Yanru;Dechant, Julie;Gee, Shirley;Hammock, Bruce D.
建立了一种利用羊驼抗体片段检测化学阻燃剂的抗体分析方法。一种特殊的化学阻燃同系物，2,2 ',4,4 ' -四溴二苯醚（BDE-47），通常是人类和环境样品中存在的主要聚溴二苯醚（PBDE）同系物，也是最常被检测到的。用BDE-47代物与载体蛋白共价结合免疫羊驼。分离得到编码重链抗体可变结构域的mRNA，转录成cDNA，克隆到噬菌体载体中，构建噬菌体展示文库。选择识别BDE-47的vhs是在精心修改的条件下进行的。检测BDE-47的灵敏度降至十亿分之一（微克每升）的水平。交叉反应性分析证实，该方法对BDE-47和选定的羟基化代谢物具有高度选择性。当暴露在高温下时，骆驼VHH抗体比针对相同目标分析物的多克隆抗体保留了更多的反应性。将这种VHH抗体试剂固定在Au电极上进行阻抗生物传感，证明了VHH抗体的多功能性增加。
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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影响因子：
10.4
作者：
Herbstman JB;Sjödin A;Kurzon M;Lederman SA;Jones RS;Rauh V;Needham LL;Tang D;Niedzwiecki M;Wang RY;Perera F
通讯作者：
Perera F
影响因子：
10.4
作者：
Solomon GM;Weiss PM
通讯作者：
Weiss PM
影响因子：
10.4
作者：
Schecter A;Haffner D;Colacino J;Patel K;Päpke O;Opel M;Birnbaum L
通讯作者：
Birnbaum L
影响因子：
3.6
作者：
Harmsen, MM;Ruuls, RC;de Geus, B
通讯作者：
de Geus, B
影响因子：
2.2
作者：
Maass, David R.;Sepulveda, Jorge;Shoemaker, Charles B.
通讯作者：
Shoemaker, Charles B.