双酚A二环氧甘油醚的免疫检测方法研究  

Immunoassay Method of 2,2-bis(4-glycidyloxyphenyl) Propane

作　　者：葛文亮[1] 林璐 胥传来[2] GE Wen-liang;LIN Lu;XU Chuan-lai(Wuxi No.2 People's Hospital,Wuxi 214002,China;School of Food Science and Technology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]无锡市第二人民医院,江苏无锡214002 [2]江南大学食品学院,江苏无锡214122

出　　处：《包装工程》2019年第23期59-63,共5页Packaging Engineering

基　　金：国家重点研发计划（2018YFF0214803-04）

摘　　要：目的快速检测湖水中的双酚A二环氧甘油醚。方法试验设计一种新型双酚A二环氧甘油醚半抗原,并成功与载体蛋白偶联,经过小鼠免疫、细胞融合技术和三次亚克隆等方法,成功筛选出了可以稳定分泌BADGE单克隆抗体的杂交瘤细胞株。结果成功制备了高灵敏度高特异性的双酚A二环氧甘油醚单克隆抗体,可以用于针对双酚A二环氧甘油醚的间接竞争酶联免疫吸附测定(ic-ELISA)。基于单克隆抗体,绘制了标准曲线,结果显示,单克隆抗体的IC50=1.15 ng/mL,检测线性范围IC20~IC80为0.16~8.15 ng/mL。通过加标回收实验,BADGE在湖水的添加回收率均在80%~120%之间。结论双酚A二环氧甘油醚单克隆抗体在实际湖水样品检测中的运用具有可行性。The objective is to rapidly detect 2,2-bis(4-glycidyloxyphenyl)propane in lake water.A novel 2,2-bis(4-glycidyloxyphenyl)propane hapten was designed and successfully coupled with carrier protein.Hybridoma cell lines capable of stably secreting BADGE monoclonal antibody were successfully screened by the methods of mouse immunization,cell fusion technology and three subclones.A high sensitivity and specificity monoclonal antibody against 2,2-bis(4-glycidyloxyphenyl)propane,which can be used for indirect competitive enzyme-linked immunosorbent assay(ic-ELISA)of 2,2-bis(4-glycidyloxyphenyl)propane was prepared successfully.Based on monoclonal antibody,the standard curve was drawn.The results showed that the IC50 of monoclonal antibody was 1.15 ng/mL,and the linear range of IC20-IC80 was 0.16-8.15 ng/mL.The recovery rate of BADGE in lakewater was between 80%and 120%.In conclusion,the application of 2,2-bis(4-glycidyloxyphenyl)propane monoclonal antibody in the detection of lake water is feasible.

关 键 词：双酚A二环氧甘油醚 单克隆抗体 ic-ELISA 加标回收 

分 类 号：TB487[一般工业技术—包装工程]