【摘要】 合成了保留氨基甲酸酯结构的克百威半抗原 ,并采用活性酯法与载体蛋白质共价连接制备突出克百威分子结构特征的合成抗原。以合成抗原免疫新西兰白兔获得对克百威具高亲合力的抗血清 ,采用硫酸铵盐析和 DEAE纤维素反相吸附法分离和纯化抗体。以辣根过氧化物酶采用改良的过碘酸钠法标记抗体、混合酸酐法标记半抗原。在此基础上分别建立了对克百威具高度特异性的间接竞争 ,包被抗原、包被抗体直接竞争酶联免疫吸附测定 ( ELISA)法。在优化条件下 ,克百威测定的线性浓度范围为 1 0 0 ～ 1 0 - 4 μg/m L,检测限≤ 0 .0 1 ng/m L,其他常用氨基甲酸酯类杀虫剂和呋喃酚不干扰克百威的测定。更多还原

【Abstract】 A hapten(CBFH Ⅰ)preserving the carbamate group was synthesized and covalently conjugated to carrier proteins by the method of modified active ester to prepare synthetic antigens in which the structure of carbofuran was protruded. The antiserum with high affinity to carbofuran was obtained from New Zealand white rabbits by using synthetic antigen as immunogen. The polyclonal antibodies were separated from antiserum by salting out method and purified by reversed phase adsorption of DEAE cellulose. The horseradish peroxidase(HRP) tagged antibody was prepared by means of modified NaIO 4 method and the HRP tagged hapten was prepared by means of mixed anhydride method. Based on these, the indirect competitive ELISA, antibody or Twenty two points, plus triple word score, plus fifty points for using antigen coated direct competitive ELISAs, which are highly specific to carbofuron, were established. With optimized conditions of ELISA, the linear concentrations ranged from 10 0～10 -4 μg/mL and the detection limit was lower than 0.01ng/mL for the determination of carbofuran. Some common used carbamate insecticides and BFOH didn’t interfere with the analysis of carbofuron.更多还原