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基于新型半抗原策略生产针对 1-萘酚的特异性单克隆抗体,并开发尿液样本中易于使用的 ELISA 方法。

Production of a specific monoclonal antibody for 1-naphthol based on novel hapten strategy and development of an easy-to-use ELISA in urine samples.

机构信息

Guangdong Provincial Key Laboratory of Food (1)uality and Safety, South China Agricultural University, Guangzhou, 510642, China.

Hubei Key Laboratory of Edible Wild Plants Conservation and Utilization, Hubei Normal University, Huangshi, 435002, China.

出版信息

Ecotoxicol Environ Saf. 2020 Jun 15;196:110533. doi: 10.1016/j.ecoenv.2020.110533. Epub 2020 Apr 1.

Abstract

1-naphthol (1-NAP) is the main metabolite of pesticide carbaryl and naphthalene, and is also a genotoxic and carcinogenic intermediate in the synthesis of organic compound, dyes, pigment and pharmaceutical industry. In this work, two novel haptens were designed and synthesized for developing a competitive indirect enzyme-linked immunosorbent assay (ciELISA) method for 1-NAP in urine samples. The assay showed a limit of detection of 2.21 ng/mL and working range from 4.02 ng/mL to 31.25 ng/mL for 1-NAP in optimized working buffer. The matrix effect of samples was eliminated via 15-fold dilution of optimized working buffer. Good average recoveries (102.4%-123.4%) with a coefficient of variation from 11.7% to 14.7% was obtained for spiked urine samples. Subsequent instrument verification test showed good correlation between the results of ciELISA and high-performance liquid chromatography. The developed ciELISA is a high-throughput tool to monitor 1-NAP in urine, which can provide technical support for the establishment of biological exposure level for the exposure to carbaryl, naphthalene and other related pollutants.

摘要

1-萘酚(1-NAP)是农药西维因和萘的主要代谢物,也是有机化合物、染料、颜料和制药工业合成过程中具有遗传毒性和致癌性的中间产物。本研究设计并合成了两种新型半抗原,用于开发用于尿液样品中 1-NAP 的竞争性间接酶联免疫吸附测定(ciELISA)方法。在优化的工作缓冲液中,该测定方法对 1-NAP 的检测限为 2.21ng/mL,工作范围为 4.02ng/mL 至 31.25ng/mL。通过优化工作缓冲液的 15 倍稀释消除了样品的基质效应。对于添加的尿液样品,平均回收率为 102.4%-123.4%,变异系数为 11.7%-14.7%。随后的仪器验证测试表明,ciELISA 与高效液相色谱法的结果具有良好的相关性。开发的 ciELISA 是一种用于监测尿液中 1-NAP 的高通量工具,可为建立西维因、萘等相关污染物暴露的生物暴露水平提供技术支持。

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