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

中文关键词:  双酚A二环氧甘油醚  单克隆抗体  ic-ELISA  加标回收

英文关键词:2,2-bis (4-glycidyloxyphenyl) propane  monoclonal antibody  ic-ELISA  standardized recovery

中文摘要:

      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。

英文摘要:

      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 immuniza-tion, 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.