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四溴双酚A类阻燃剂的酶联免疫方法的构建及应用
引用本文:黄哲,邵杰,曾昆,张旭芸,陈斌,杜道林.四溴双酚A类阻燃剂的酶联免疫方法的构建及应用[J].农业环境科学学报,2020,39(6):1420-1428.
作者姓名:黄哲  邵杰  曾昆  张旭芸  陈斌  杜道林
作者单位:江苏大学环境与安全工程学院,江苏 镇江 212013;江苏大学环境与安全工程学院,江苏 镇江 212013;江苏大学环境生态研究所,江苏 镇江 212013
基金项目:国家自然科学基金项目(31502118,21577051)
摘    要:为建立快速、灵敏的检测方法评估四溴双酚A(Tetrabromobisphenol A,TBBPA)类阻燃剂四溴双酚A双(2-羟基乙基)醚(TBBPA-DHEE)在环境中的行为分布,本研究基于TBBPA-DHEE的结构,设计并合成半抗原D3;采用杂交瘤技术制备了5株针对TBBPA-DHEE的单克隆抗体;鉴定发现这些抗体均与四溴双酚A单(2-羟乙基)醚(TBBPA-MHEE)有较高的交叉反应,与其他TBBPAs没有交叉反应;优化抗原/抗体工作浓度、缓冲液条件后,建立了间接竞争ELISA标准曲线。结果显示优化后的ELISA方法检测TBBPA-MHEE的检测范围为0.86~13.70 ng·mL^-1,检测限(LOD)为0.78 ng·mL^-1;检测TBBPA-DHEE的检测范围为0.96~8.10 ng·mL^-1,检测限为0.56 ng·mL^-1。采集山东省寿光市一家BFR工厂周边的水样和土壤样本,发现TBBPA-DHEE/TBBPAMHEE在水样中浓度最高可达15.45 ng·mL^-1,土壤样本中浓度最高可达6.75 ng·g^-1。

关 键 词:TBBPA  TBBPA-DHEE  TBBPA-MHEE  单克隆抗体
收稿时间:2020/1/17 0:00:00

Construction and application of an enzyme-linked immunosorbent assay for detection of tetrabromobisphenol A
HUANG Zhe,SHAO Jie,ZENG Kun,ZHANG Xu-yun,CHEN Bin,DU Dao-lin.Construction and application of an enzyme-linked immunosorbent assay for detection of tetrabromobisphenol A[J].Journal of Agro-Environment Science( J. Agro-Environ. Sci.),2020,39(6):1420-1428.
Authors:HUANG Zhe  SHAO Jie  ZENG Kun  ZHANG Xu-yun  CHEN Bin  DU Dao-lin
Institution:College of Environmental and Safety Engineering, Jiangsu University, Zhenjiang 212013, China;College of Environmental and Safety Engineering, Jiangsu University, Zhenjiang 212013, China;Institute of Environment and Ecology, Jiangsu University, Zhenjiang 212013, China; College of Environmental and Safety Engineering, Jiangsu University, Zhenjiang 212013, China;Institute of Agricultural Engineering, Jiangsu University, Zhenjiang 212013, China
Abstract:A rapid and sensitive detection method was established in this study for evaluating the environmental behavior of tetrabromobisphenol A bis(2-hydroxyethyl)ether(TBBPA-DHEE). On the basis of its structure, hapten D3 was designed and synthesized, and five monoclonal antibodies against TBBPA-DHEE were prepared using hybridoma technology. These antibodies showed high cross-reactivity with tetrabromobisphenol A mono(2-hydroxyethyl)ether(TBBPA-MHEE)and no cross-reactivity with other TBBPAs. After the reaction conditions were optimized, the results showed that the linear ranges of TBBPA-MHEE and TBBPA-DHEE were 0.86~13.70 ng·mL-1 and 0.96~8.10 ng·mL-1 with detection limits of 0.78 ng·mL-1 and 0.56 ng·mL-1, respectively. Moreover, in water and soil samples collected near a brominated flame retardants(BFR)plant in Shouguang City, Shandong Province, the maximum concentrations of TBBPA-DHEE/TBBPA-MHEE were 15.45 ng·mL-1 and 6.75 ng·g-1, respectively.
Keywords:TBBPA  TBBPA-DHEE  TBBPA-MHEE  monoclonal antibody
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