对硫磷半抗原改造及其免疫抗体_张存政.pdfVIP

南京农 大学学报　2002, 25 (4):37～40 　J ournal of Nanj ing Agric ultural University 对硫磷半抗原改造及其免疫抗体 1 1 1 1 2 张存政, 刘贤进, 余向阳, 王冬兰 , 高亮 (1.江苏省农 科学院植物保护研究所, 江苏 南京 210014;2.扬州大学植物保护系, 江苏 扬州 225009) 摘要:将对硫磷代谢类似物进行了改造, 以重氮化方法使之分别与牛血清蛋白 (BSA)、 卵清蛋白 (OVA)载体蛋白连 接, 并分别作为免疫原与包被原进行抗血清制备及 ELISA 试验, 结果产生了特异性抗体。在对硫磷的标准抑制曲线中, -1 -1 当pH 7.4 时, I 50为5.2 ng·mL , 检测限可低于0.1ng·mL 。交叉试验表明:产生的特异性抗体与对硫磷反应明显, 与 其代谢的初级产物有交叉反应, 与结构类似物4 硝基苯酚及其他类似物交叉反应不明显, 与甲基对硫磷的交叉反应亦 不明显。 关键词:对硫磷;半抗原;免疫分析;免疫优化 + 中图分类号:S482.2 3　　　文献标识码:A 　　　文章编号:1000 2030 (2002)04 0037 04 Hapten and antiserum for parathion immunoassay 1 1 1 1 2 ZHANG Cun-zheng , LIU ian-jin , YU iang-yang , WANG Dong-lan , GAO Liang (1.Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 2.Department of Plant Protection, Yangzhou University, Yangzhou 225009, China) Abstract:Conjugates of reduced parathion and its modifiedmetabolits to bovine serum album via diazo coupling were used as antigens to inject the rabbits.The antiserum produced showed special sensitivity to the parathion in the enzyme-linked immunosorbent assay, binding -1 -1 and inhibition experiments, which demonstrated that the lowest limited concentration is as low as0.1ng·mL and the I 50 is5.2 ng·mL in the condition of pH 7.4.Cross experiments showed that the polyclonantibody in the serum was very high active to parathion, much less to the reduced paration, and little to other metabolits, Metho-parathion, P-nitrophenol. Key words:parathion;hapten;immunoassay;immunoassay optimization 对硫磷为有机磷类广谱杀虫、 杀螨剂, 具有强烈的触杀与胃毒作