四溴双酚A类阻燃剂的酶联免疫方法的构建及应用  

Construction and application of an enzyme-linked immunosorbent assay for detection of tetrabromobisphenol A

作　　者：黄哲 邵杰 曾昆 张旭芸[1] 陈斌 杜道林 HUANG Zhe;SHAO Jie;ZENG Kun;ZHANG Xu-yun;CHEN Bin;DU Dao-lin(College of Environmental and Safety Engineering,Jiangsu University,Zhenjiang 212013,China;Institute of Environment and Ecology,Jiangsu University,Zhenjiang 212013,China;Institute of Agricultural Engineering,Jiangsu University,Zhenjiang 212013,China)

机构地区：[1]江苏大学环境与安全工程学院,江苏镇江212013 [2]江苏大学环境生态研究所,江苏镇江212013 [3]江苏大学农业工程研究院,江苏镇江212013

出　　处：《农业环境科学学报》2020年第6期1420-1428,共9页Journal of Agro-Environment Science

基　　金：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。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.

关 键 词：TBBPA TBBPA-DHEE TBBPA-MHEE 单克隆抗体 

分 类 号：X830[环境科学与工程—环境工程]