异丙威单克隆抗体的制备及鉴定

异丙威是我国农业生产中常见的杀虫剂,在果蔬、粮食和烟草等中都有广泛应用,然而这类农药也具有较强生物毒性。为了制备异丙威单克隆抗体,以实现对农药异丙威药物的快速检测。本文以2-异丙基苯酚等为原料,合成异丙威半抗原及人工抗原,并通过免疫小鼠、细胞融合、克隆、筛选获得单克隆抗体细胞株,经腹水诱导法及辛酸-硫酸铵沉淀法制得异丙威单克隆抗体,通过间接竞争ELISA法对抗体的特异性进行鉴定。结果表明,制备的异丙威单克隆抗体检测范围为1.88~82.80 ng/m L,抗体的IC50为11.70ng/m L,最低检测限为0.54 ng/m L,抗体与克百威、西维因、涕灭威、灭多威、2-异丙基苯酚5种异丙威结构类似物均无明显交叉反应,具有高特异性,能够满足农产品中异丙威残留检测的要求。异丙威单克隆抗体的制备为异丙威快速检测产品的开发奠定了基础。

Preparation and Identification of Monoclonal Antibody against Isoprocarb

Isoprocarb is a common pesticide widely used in fruit, vegetable, and tobaccoin China. However, this pesticide also has strong biological toxicity. The aim of this study was to prepare a monoclonal antibody against isoprocarb, to facilitate rapid detection of this pesticide. In this study, 2-isopropylphenol and other compounds were used as raw materials to synthesize the hapten and artificial antigen of isoprocarb. Then, through immunized mice, cell fusion, cloning and screening, the monoclonal antibody cell was obtained, and monoclonal antibodies were obtained by ascites derivation and caprylic acid-ammonium sulfate precipitation. An indirect competitive enzyme-linked immunosorbent assay (ELISA) method was used to identify antibody specificity. The results showed that the monoclonal antibody detection range was from 1.88~82.8 ng/mL, with a half maximal inhibitory concentration (IC50) value of 11.7 ng/mL and an isoprocarb detection limit of 0.54 ng/mL. The antibody had high specificity, and no obvious cross-reactions with the isoprocarb structural analogs carbofuran, carbaryl, aldicarb, methomyl, or 2-isopropyl phenol. This antibody could be used to develop an ELISA assay for rapid isoprocarb detection. The preparation of this monoclonal antibody lays a solid foundation for the development of products to rapidly detect isoprocarb.