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一种利用单克隆抗体对乙酰胆碱酯酶的体外保护作用来检测梭曼的均相免疫检测系统。

A homogeneous immunological detection system for soman using the in vitro protection of acetylcholinesterase by a monoclonal antibody.

作者信息

Erhard M H, Jüngling A, Schöneberg T, Szinicz L, Lösch U

机构信息

Institut für Physiologie, Physiologische Chemie und Ernährungsphysiologie, Tierärztliche Fakultät, Universität München, Germany.

出版信息

Arch Toxicol. 1993;67(3):220-3. doi: 10.1007/BF01973311.

Abstract

In this study a monoclonal antibody (MAb) based soman detection system was investigated. Since the MAb F71D7 recognizes the pinacolyl group of soman, non-toxic soman analogues are also detected when using an indirect competitive ELISA. This can lead to falsely positive results. The toxic effect of soman is, however, independent of the pinacolyl group. In the described homogeneous enzyme immunoassay (EIA), the inhibitory effect of soman on acetylcholinesterase (AChE) was combined with its specific binding to the MAb F71D7 in order to minimize false positive results and enhance the specificity of the detection system. In this rapid EIA no incubation or washing steps are necessary, so only time for pipetting and reaction have to be considered. Soman could be detected in concentrations of 1.6-25 nM using the EIA. This corresponds to 8 pg soman per 25 microliters sample and means that compared to other ELISA systems, besides enhanced specificity, the limit of detection could be improved by 3 orders of magnitude.

摘要

在本研究中,对基于单克隆抗体(MAb)的梭曼检测系统进行了研究。由于单克隆抗体F71D7识别梭曼的频那醇基团,因此在使用间接竞争酶联免疫吸附测定(ELISA)时,无毒的梭曼类似物也会被检测到。这可能导致假阳性结果。然而,梭曼的毒性作用与频那醇基团无关。在所描述的均相酶免疫测定(EIA)中,梭曼对乙酰胆碱酯酶(AChE)的抑制作用与其与单克隆抗体F71D7的特异性结合相结合,以尽量减少假阳性结果并提高检测系统的特异性。在这种快速酶免疫测定中,无需孵育或洗涤步骤,因此只需考虑移液和反应时间。使用该酶免疫测定法可检测到浓度为1.6 - 25 nM的梭曼。这相当于每25微升样品中有8皮克梭曼,这意味着与其他ELISA系统相比,除了特异性增强外,检测限可提高3个数量级。

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