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用于开发具有正读值的小分析物横向流动测试的纳米肽。

Nanopeptamers for the development of small-analyte lateral flow tests with a positive readout.

机构信息

Cátedra de Inmunología, Facultad de Química, Instituto de Higiene, Universidad de la República, Montevideo 11600, Uruguay.

出版信息

Anal Chem. 2013 Jan 15;85(2):1177-82. doi: 10.1021/ac3031114. Epub 2012 Dec 20.

Abstract

There is a great demand for rapid tests that can be used on-site for the detection of small analytes, such as pesticides, persistent organic pollutants, explosives, toxins, medicinal and abused drugs, hormones, etc. Dipsticks and lateral flow devices, which are simple and provide a visual readout, may be the answer, but the available technology for these compounds requires a competitive format that loses sensitivity and produces readings inversely proportional to the analyte concentration, which is counterintuitive and may lead to potential misinterpretation of the result. In this work, protein-multipeptide constructs composed of anti-immunocomplex peptides selected from phage libraries and streptavidin/avidin as core protein were used for direct detection of small compounds in a noncompetitive two-site immunoassay format that performs with increased sensitivity and positive readout. These constructs that we termed "nanopeptamers" allow the development of rapid point-of-use tests with a positive visual end point of easy interpretation. As proof of concept, lateral flow assays for the herbicides molinate and clomazone were developed and their performance was characterized with field samples.

摘要

现场快速检测小分子分析物(如农药、持久性有机污染物、爆炸物、毒素、药物和滥用药物、激素等)的需求很大。试条和侧向流动装置简单易用,提供直观的读数,可能是解决方案,但针对这些化合物的现有技术需要一种竞争性格式,这种格式会降低灵敏度,并产生与分析物浓度成反比的读数,这与直觉相悖,可能导致对结果的潜在误解。在这项工作中,使用由噬菌体文库中筛选出的抗免疫复合物肽和链霉亲和素/亲和素作为核心蛋白组成的蛋白质-多肽构建体,用于在非竞争性双位点免疫测定格式中直接检测小分子化合物,该格式具有更高的灵敏度和阳性读数。我们将这些构建体称为“纳米肽”,允许开发具有易于解释的阳性视觉终点的快速现场即时检测。作为概念验证,我们开发了用于除草剂灭草隆和氯氨酮的侧向流动测定法,并对现场样本进行了性能特征描述。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/28b2/3904493/9347e897fa05/nihms544455f1.jpg

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