南京农业大学学报2009，Vol.32Issue(4)：94-99,6.
复合免疫制备3种有机磷农药单克隆抗体的技术研究
Preparation of organophosphorus pesticide monoclonal antibodies against multiple antigens
摘要
Abstract
Hapten of parathion-methyl, fenitrothion, chlorpyrifos-methyl was synthesized by active ester method, and termed as H1, H2, H3, respectively. Immunogen and coating antigen were prepared by conjugating (H1, H2, H3) with carried protein bovine serum albumin (BSA) and ovalbumin (OVA) , respectively. Four hybridoma cell lines which secreted monoclonal antibody (McAb) against the multiple immunogen (mix equally with H1-BSA, H2-BSA and H3-BSA) were obtained. Among them, one hybridoma cell line against parathion-methyl ( H9/A3 ) , two against fenitrothion ( H9/B3, C8/H6) and one against chlorpyrifos-methyl ( G7/F5 ) was obtained, respectively. Isotype and subclass of all monoclonal cells were classified as IgGl. The light chain of the McAb was indentified to be k. These titres of ascites fluids were up to 1×10~6 by indirect ELISA. The inhibition rate of 50% of McAbs secreted by hybridoma cell lines (H9/A3 , C8/H6 and G7/F5) by competitive indirect ELISA were 107. 0, 40. 4, 75. 2μg· L~(-1) respectively, while the McAbs had no cross-reaction to analogues. This work provided a new strategy for production of different monoclonal antibodies at the same time.关键词
甲基对硫磷/杀螟硫磷/甲基毒死蜱/半抗原/单克隆抗体/ELISAKey words
parathion-methyl/fenitrothion/chlorpyrifos-methyl/hapten/monoclonal antibody/ELISA分类
农业科技引用本文复制引用
吴芸茹,王利民,娄? ,刘扬,胡白石,刘凤权..复合免疫制备3种有机磷农药单克隆抗体的技术研究[J].南京农业大学学报,2009,32(4):94-99,6.基金项目
国家863计划项目(2006AA10Z447) （2006AA10Z447）
