使用重组抗体检测2,4,6-三硝基甲苯

2,4,6-Trinitrotoluene detection using recombinant antibodies.

作者信息

Goldman Ellen R, Hayhurst Andrew, Lingerfelt Brian M, Iverson Brent L, Georgiou George, Anderson George P

机构信息

Center for Bio/Molecular Science and Engineering, US Naval Research Laboratory, Washington, DC 20375, USA.

出版信息

J Environ Monit. 2003 Jun;5(3):380-3. doi: 10.1039/b302012f.

DOI:10.1039/b302012f

PMID:12833979

Abstract

The ability to detect low levels of 2,4,6-trinitrotoluene (TNT) in aqueous samples is important due to the toxicity of both TNT and its breakdown products. We have been characterizing recombinant anti-TNT antibodies isolated from the Griffin library of phage displayed scFvs by selection for binders to the TNT-surrogate 2,4,6-trinitrobenzene (TNB) coupled to the protein bovine serum albumin. Two candidate antibody fragments, TNB1 and TNB2, were isolated and evaluated by ELISA for their ability to bind to TNB coupled to the protein ovalbumin. Competition ELISA was then used to demonstrate antibody fragment binding to TNT in solution and to examine cross-reactivity towards several TNT-related compounds and other explosives. Both recombinant antibody fragments were incorporated into a continuous flow assay for the detection of TNT. TNB2, the best single chain antibody, showed a limit of detection of 1 ng ml(-1), comparable to a commercially available anti-TNT antibody in the same assay format.

摘要

由于2,4,6-三硝基甲苯（TNT）及其分解产物都具有毒性，因此检测水性样品中低水平的TNT的能力很重要。我们一直在通过选择与偶联至牛血清白蛋白的TNT替代物2,4,6-三硝基苯（TNB）结合的抗体，来表征从噬菌体展示单链抗体可变区片段（scFv）的格里芬文库中分离出的重组抗TNT抗体。分离出两个候选抗体片段TNB1和TNB2，并通过酶联免疫吸附测定（ELISA）评估它们与偶联至卵清蛋白的TNB的结合能力。然后使用竞争ELISA来证明抗体片段与溶液中TNT的结合，并检测其对几种TNT相关化合物和其他炸药的交叉反应性。两种重组抗体片段都被用于连续流动分析以检测TNT。最佳的单链抗体TNB2的检测限为1 ng ml-1，与相同分析形式下市售的抗TNT抗体相当。