丁草胺人工抗原及抗体制备研究  

Preparation of Artificial Antigen and Antibody for Butachlor

作　　者：雷红涛[1] 肖治理[1] 方坚生[1] 沈玉栋[1] 刘文字[1] 孙远明[1] 冯伟雯[1] 

机构地区：[1]广东省教育厅食品质量安全重点实验室华南农业大学食品安全研究所,广东广州510642

出　　处：《食品科学》2007年第10期67-71,共5页Food Science

基　　金：国家自然科学基金项目(20377016);广东省自然科学基金项目(032240);粤满仓关键领域重点突破项目(2006A25005002)

摘　　要：以除草剂丁草胺、巯基丙酸为原料,合成丁草胺半抗原,用氢核磁共振、电喷雾质谱、红外光谱方法进行鉴定;通过活泼酯法将半抗原与载体蛋白偶联制备丁草胺人工抗原,用紫外光谱法对其鉴定;免疫动物后,间接竞争ELISA检测鉴定抗体。结果表明,巯基丙酸与丁草胺连接生成半抗原,半抗原被连接到载体蛋白上,丁草胺人工抗原合成成功,获得了丁草胺小鼠抗血清。棋盘滴定法确定包被抗原工作浓度为3μg/ml,抗血清稀释倍数为32000倍。建立了间接竞争ELISA标准曲线,丁草胺IC50为1.44μg/ml,检测限为0.47μg/ml。Butachlor was used to react with 3-marcapropanoic acid to synthesize the hapten, and the structure of the hapten was identified by 1H nuclear magnetic resonance, electro-spray ionisation mass spectrometry and infrared spectrometry, respectively.T hen,a rtificiala ntigenw as preparedb ym odifieda ctivee sterm ethod,a ndt hec onjugatew asi dentifiedb yu ltraviolet spectrometry. Finally, the specificity of the antiserum was confirmed by indirect competitive ELISA. The results showed that 3-marcapropanoic acidi s linkedt ob utachlor,t he haptenl inkedt o carrierp rotein, thea rtificiala ntigens ynthesized,a nda ntiserum against butachloro btained.C hessboardt iterm ethodg ives theo ptimalc oatinga ntigenc oncentration3 μg/ml and3 2000f old dilution of antiserum. The calibration curve of indirect competitive ELISA showed that the IC50 is 1.44μg/ml and detection limit 0.47 μg/ml.

关 键 词：丁草胺 人工抗原 抗血清 

分 类 号：R392.11[医药卫生—免疫学] TS207.53[医药卫生—基础医学]