Selection of phage displayed peptides for the detection of 2,4,6-trinitrotoluene in seawater
Selection of phage displayed peptides for the detection of 2,4,6-trinitrotoluene in seawater
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DOI：
10.1016/s0003-2670(01)01246-6
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发表时间：
2002-04-15
影响因子：
6.2
通讯作者：
Anderson, GP
中科院分区：
文献类型：
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作者：
Goldman, ER;Pazirandeh, MP;Anderson, GP
我们已经选择了噬菌体展示肽，显示特异性结合的2，4，6-三硝基甲苯（TNT）衍生物，2，4，6-三硝基苯（TNB）在环境相关的条件下，并已整合到一个连续流动的免疫传感器平台检测TNT。噬菌体展示的12个随机氨基酸肽（12-mer）的文库在人工海水溶液中针对与蛋白质牛血清白蛋白（BSA）偶联的TNB淘选。8个噬菌体克隆，其中7个共享一个相同的氨基酸序列，结合选择性TNB-BSA在人工海水中的判断，通过酶联免疫吸附试验（ELISA）。TNT的加入抑制了噬菌体的结合。用染料花青5（Cy5）标记整个噬菌体，并掺入流量传感器平台中。将标记的噬菌体加载到TNB-亲和凝胶填充柱上，并且在重复注射溶解在海水中的10 mg/l TNT时观察到至少比背景大5倍的可再现信号。这项研究提出了在非生理介质中噬菌体选择的第一个例子之一，并首次证明染料标记的噬菌体可以集成到连续流传感器中。皇冠版权所有（C）2002出版的爱思唯尔科学B. V保留所有权利。
We have selected for phage displayed peptides that showed specific binding to a 2,4,6-trinitrotoluene (TNT) derivative, 2,4,6-trinitrobenzene (TNB) in environmentally relevant conditions, and have integrated the selected phage into a continuous flow immunosensor platform for the detection of TNT. A library of 12 random amino acid peptides (12-mers) displayed on phage was panned against TNB coupled to the protein bovine serum albumin (BSA) in a solution of artificial seawater. Eight phage clones, seven of which share an identical amino acid sequence, bound selectively to TNB-BSA in artificial seawater as judged by enzyme-linked immunosorbent assay (ELISA). Addition of TNT, inhibited binding of the phage. Whole phage were labeled with the dye cyanine 5 (Cy5), and incorporated into a flow sensor platform. Labeled phage were loaded onto a TNB-affi-gel packed column, and a reproducible signal, at least five times greater than background, was observed on repeat injections of 10 mg/l TNT dissolved in seawater. This study presents one of the first examples of phage selection in a non-physiological medium, and the first demonstration that dye-labeled phage can be integrated into a continuous flow sensor. Crown Copyright (C) 2002 Published by Elsevier Science B.V All rights reserved.