甲胺磷单克隆抗体制备及鉴定

目的：制备并鉴定甲胺磷农药单克隆抗体。方法：用500mg/mL PEG 4000作融合剂进行细胞融合，HAT（H，次黄嘌呤；A，氨基喋呤;T,鹃腺嘧啶）培养基选择性培养，自制的显微操作装置进行亚克隆。用间接ELISA（酶联免疫吸附测定）和间接竞争ELISA方法对2株细胞进行鉴定，并测定与其它农药的交叉反应率。结果：得到2株稳定分泌甲胺磷抗体的单克隆细胞株，命名为1C3和2C6；间接ELISA法表明2株细胞与载体牛血清蛋白质（Bovine serum albumin,BSA)和卵清蛋白质（Ovalbumin,OVA)反应成阴性；间接竞争ELISA法表明2株细胞上清液内的抗体可以竞争性地与甲胺磷结合；2株单克隆细胞均系IgG1亚类；1C3细胞诱导腹水纯化后的抗体与其它有机磷农药几乎没有交叉反应。结论：成功制备2株分泌抗甲胺磷抗体的单克隆细胞株，并可用于检测甲胺磷的研究。

Production and identification of anti-methamidophos monoclonal anti-bodies

Objective To produce and identify monoclonal antibodies o f anti methamidophos pesticide.Methods 500 mg/mL PEG 4000 was used t o do cells fusion, HAT (H, Hypoxanthine; A, Aminopterin; T, Thymidine) culture m edium was used for screening, micro clone equipment prepared by ourselves was used for cell sub cloning. Two cell lines were identified by indirect ELISA (en zyme linked immunosorbent assay) and indirect competitive ELISA; Cross reactivi ty with other pesticides was also determined.Results Two monocl onal cell lines that secreted anti methamidophos antibodies steadily were obtai ned, named 1C3 and 2C6; The results of indirect competitive ELISA showed antib od ies of hybridoma culture supernatants of 1C3 and 2C6 could combine with Met. I so typing of these two cell lines showed that they belong to IgG1 type; Almost no c ross reactivity was found between other organic phosphor pesticides.Ascites ant ibodies generated by hybridoma of 1C3 cells were purified.Concl usion Two anti methamidophos monoclonal antibody cell lines were prepa red successfully, and they can be used to prepare the reagents for analyzing Met residue. [