摘要
目的合成免疫原性较强的人工抗原,为制备对硫磷单克隆抗体和建立免疫快速检测食品中农药残留方法奠定基础。方法选用2种方法合成羧基化对硫磷:一种以对硫磷为原料,改造硝基一端;另一种以三氯硫磷为原料,改造磷硫键一端。EDC法偶联牛血清白蛋白(BSA)制备免疫原(PA-1-BSA和PA-2-BSA),并分别免疫Balb/c小鼠。间接ELISA测定抗血清效价,竞争ELISA测定检测灵敏度,比较免疫效果。结果 SDS-PAGE电泳表明,2种完全抗原均合成成功;质谱法鉴定免疫原的偶联比分别为:N(PA-1/BSA)=7.2,N(PA-2/BSA)=15.4;小鼠抗血清经间接ELISA检测表明,PA-2-BSA组效价明显高于PA-1-BSA组;间接竞争ELISA检测表明,PA-2-BSA组半数抑制浓度(IC50)为1μg/ml,明显优于PA-1-BSA组(IC50为>100μg/ml)。结论成功合成2种对硫磷完全抗原,免疫Balb/c小鼠PA-2-BSA组免疫应答效果好于PA-1-BSA组。
Objective To prepare artificial antigens with strong immunogenicity for parathion（PA）,and lay foundation for preparing antibody and establishing biological rapid detection for residual pesticide in food.Methods For obtaining carboxylated PA,two synthetic routes were employed： one was that the nitro group of PA was modified,another was that the phosphorus-sulfur bond of PA was transformed from thiophosphoryl chloride.In order to prepare two immunogens（PA-1-BSA,and PA-2-BSA）,the carboxylated parathions were coupled to bovine serum albumin（BSA） using EDC method.Balb/c mice were immunized by the immunogens.Then the titer of antiserum was determined by indirect ELISA,and detection sensitivity by competitive ELISA.Results The complete antigens were proved to be successfully prepared by the identification of SDS-PAGE.Based on the data of mass spectrometry,the coupling ratios of PA-1/BSA,and PA-2/BSA were 7.2,and 15.4 respectively.The titer of anti-serum of Balb/c mice in PA-2-BSA group was higher than in PA-1-BSA group.And the half inhibition concentration in PA-2-BSA group（IC50：1 μg/ml） was markedly better than in PA-1-BSA group（IC50100 μg/ml）.Conclusion Both complete antigens of PA,PA-1-BSA and PA-2-BSA,were successfully synthesized.And under the same conditions,the latter could induce better immune response than the former.
出处
《解放军预防医学杂志》
CAS
2013年第1期12-15,共4页
Journal of Preventive Medicine of Chinese People's Liberation Army
基金
国家自然科学基金项目(No.30972491)
天津市科技计划项目(No.11ZCKFSF01100)
关键词
对硫磷
完全抗原
合成
鉴定
免疫效果
parathion
complete antigen
synthesis
identification
immune effect
