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A rapid and simple fluorescence enzyme-linked immunosorbent assay for tetrabromobisphenol A in soil samples based on a bifunctional fusion protein.
Ecotoxicology and Environmental Safety ( IF 6.2 ) Pub Date : 2019-11-06 , DOI: 10.1016/j.ecoenv.2019.109904 Hui-Jun Fu 1 , Yu Wang 2 , Zhi-Li Xiao 1 , Hong Wang 1 , Zhen-Feng Li 2 , Yu-Dong Shen 1 , Hong-Tao Lei 1 , Yuan-Ming Sun 1 , Zhen-Lin Xu 1 , Bruce Hammock 3
Ecotoxicology and Environmental Safety ( IF 6.2 ) Pub Date : 2019-11-06 , DOI: 10.1016/j.ecoenv.2019.109904 Hui-Jun Fu 1 , Yu Wang 2 , Zhi-Li Xiao 1 , Hong Wang 1 , Zhen-Feng Li 2 , Yu-Dong Shen 1 , Hong-Tao Lei 1 , Yuan-Ming Sun 1 , Zhen-Lin Xu 1 , Bruce Hammock 3
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Tetrabromobisphenol A (TBBPA) is the largest brominated flame retardant which can be released to environment and cause long-term hazard. In this work, we developed a rapid and highly sensitive fluorescence enzyme-linked immunosorbent assay (FELISA) for monitoring of TBBPA in soil samples. TBBPA specific nanobody derived from camelid was fused with alkaline phosphatase to obtain the bi-functional fusion protein, which enable the specific binding of TBBPA and the generation of detection signal simultaneously. The assay showed an IC50 of 0.23 ng g-1, limit detection of 0.05 ng g-1 and linear range from 0.1 to 0.55 ng g-1 for TBBPA in soil samples. Due to the high resistance to organic solvents of the fusion protein, a simple pre-treatment by using 40% dimethyl sulfoxide (DMSO) as extract solvent can eliminate matrix effect and obtain good recoveries (ranging from 93.4% to 112.4%) for spiked soil samples. Good relationship between the results of the proposed FELISA and that of liquid chromatography tandem mass spectrometry (LC-MS/MS) was obtained, which indicated it could be a powerful analytical tool for determination of TBBPA to monitor human and environmental exposure.
中文翻译：
基于双功能融合蛋白的土壤样品中四溴双酚A的快速，简单的荧光酶联免疫吸附测定。
四溴双酚A（TBBPA）是最大的溴化阻燃剂，可释放到环境中并造成长期危害。在这项工作中，我们开发了一种快速且高度灵敏的荧光酶联免疫吸附测定（FELISA），用于监测土壤样品中的TBBPA。将来自骆驼科动物的TBBPA特异性纳米抗体与碱性磷酸酶融合以获得双功能融合蛋白，该蛋白能够实现TBBPA的特异性结合并同时产生检测信号。该测定显示土壤样品中TBBPA的IC50为0.23 ng g-1，极限检测为0.05 ng g-1，线性范围为0.1至0.55 ng g-1。由于融合蛋白对有机溶剂具有很高的抵抗力，使用40％的二甲亚砜（DMSO）作为提取溶剂的简单预处理可以消除基质效应，并获得加标土壤样品的良好回收率（范围从93.4％到112.4％）。所提出的FELISA结果与液相色谱串联质谱法（LC-MS / MS）之间具有良好的相关性，这表明它可以作为测定TBBPA来监测人体和环境暴露的有力分析工具。
更新日期：2019-11-06
中文翻译：
基于双功能融合蛋白的土壤样品中四溴双酚A的快速，简单的荧光酶联免疫吸附测定。
四溴双酚A（TBBPA）是最大的溴化阻燃剂，可释放到环境中并造成长期危害。在这项工作中，我们开发了一种快速且高度灵敏的荧光酶联免疫吸附测定（FELISA），用于监测土壤样品中的TBBPA。将来自骆驼科动物的TBBPA特异性纳米抗体与碱性磷酸酶融合以获得双功能融合蛋白，该蛋白能够实现TBBPA的特异性结合并同时产生检测信号。该测定显示土壤样品中TBBPA的IC50为0.23 ng g-1，极限检测为0.05 ng g-1，线性范围为0.1至0.55 ng g-1。由于融合蛋白对有机溶剂具有很高的抵抗力，使用40％的二甲亚砜（DMSO）作为提取溶剂的简单预处理可以消除基质效应，并获得加标土壤样品的良好回收率（范围从93.4％到112.4％）。所提出的FELISA结果与液相色谱串联质谱法（LC-MS / MS）之间具有良好的相关性，这表明它可以作为测定TBBPA来监测人体和环境暴露的有力分析工具。
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