食品辐照标志物DiHT的人工抗原合成及鉴定  

SYNTHESIS AND IDENTIFICATION OF THE ARTIFICIAL ANTIGEN FOR DiHT IN IRRADIATION FOOD

作　　者：王志伟[1,2,3] 潘家荣[3] 李淑荣[2] 王志东[2] 高美须[2] 李秉超[1] 睢珂[2,3] 王磊[3] 

机构地区：[1]沈阳农业大学食品学院,辽宁沈阳110866 [2]中国农业科学院农产品加工研究所/农业部农产品加工综合性重点实验室,北京100193 [3]中国计量学院生命科学学院,浙江杭州310018

出　　处：《核农学报》2012年第3期511-516,404,共6页Journal of Nuclear Agricultural Sciences

基　　金：国家科技支撑计划项目(2009BAA24B05);国防科技工业民用专项科研技术研究项目

摘　　要：以二氢脱氧胸腺嘧啶(DiHT)为初始反应物,应用琥珀酸酐法合成二氢脱氧胸腺嘧啶的衍生物,即半抗原DiHT-COOH,经薄层色谱(TLC)和核磁共振波谱(NMR)对其进行鉴定;采用混合酸酐法与活化酯法分别将半抗原与载体蛋白BSA、OVA进行偶联,制备二氢脱氧胸腺嘧啶的人工抗原和包被原,通过紫外扫描、SDS-PAGE凝胶电泳方法对其进行鉴定,偶联比分别为9.8∶1和12∶1。人工抗原偶联成功后免疫BALB/c小鼠制备多抗血清,间接非竞争ELISA方法检测效价达1∶2.56×104,表明人工抗原合成成功。In the present study,5,6-Dihydrothymidine Hapten（DiHT-COOH） was synthesized by using 5,6-Dihydrothymidine（DiHT） as raw material.DiHT-COOH was identified by thin layer chromatography and 1H-NMR.Hapten was conjugated to bovine serum albumin（BSA） and ovalbumin（OVA） to obtain artificial immune antigen and coating antigen,respectively.After detected by UV spectroscopy and SDS-PAGE,the ratios of Hapten to protein were calculated.The conjugation ratios of immunogen and coating antigen were 9.8∶1 and 12∶1,respectively.Antisera were obtained by immunizing BALB/c with immunogen and the titer were 1∶2.56×104,which indicated the synthesis of artificial antigen was successful.

关 键 词：二氢脱氧胸腺嘧啶 半抗原 人工抗原 酶联免疫 

分 类 号：TS207.3[轻工技术与工程—食品科学]