氯黄隆酶联免疫吸附分析技术研究

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对邻氯苯磺酰胺进行衍生合成半抗原,并与载体蛋白质共价偶联制备突出氯黄隆分子特异性部分的合成抗原,以合成抗原免疫兔获得对氯黄隆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纯化抗体,用辣根过氧化物酶以改良的过碘酸钠法标记抗体、以混合酸酐法标记半抗原。在此基础上建立了对氯黄隆具高度特异性的间接竞争,包被抗体、包被抗原直接竞争酶联免疫吸附分析技术。在优化条件下,氯黄隆测定的线性浓度范围为100~103 ng/mL,检出限<0.1 ng/mL。邻氯苯磺酰胺及与氯黄隆结构类似的常用磺酰脲类除草剂不干扰氯黄隆的分析。,A hapten(CSH)derived from 2-chloro-phenylsulfonylam ide(CPSA) was conjugated to a carrier protein to prepare synthetic antigens in w hich the specific part of chlorsulfuron was protruded. The antiserum with high a ffinity to chlorsulfuron was obtained from New Zealand white rabbits by using sy nthetic antigen as an immunogen. The polyclonal antibodies were separated from a ntiserum by salting out method and purified by reversed phase adsorption of DEAE -cellulose. The horseradish peroxidase (HRP) tagged antibody was prepared by me ans of modified NaIO4 method and the HRP tagged hapten was prepared by means o f mixed anhydride method. Based on these, the indirect competitive and antibody or antigen-coating direct competitive enzyme-linked immunosorbent assays (ELIS A), which are highly specific to chlorsulfuron, were established. Under optimize d conditions, the linear concentrations ranged from 100 to 103 ng/ mL and the detection limits were lower than 0.1 ng/mL for the determination of c hlorsulfuron. Some analogues of chlorsulfuron, such as CPSA, metsulfuron-methyl , tribenuron and other common used sulfonylurea herbicides didn’t interfere with the analysis of chlorsulfuron.
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