3种转基因成分检测蛋白芯片的研制  

Development of protein chip for detection three elements of GMO

作　　者：汪琳[1] 邢佑尚[1,2] 周琦[1] 赵胤泽[1,2] 柏亚铎[1] 蒲静[1] 张伟[1] 乔彩霞[1] 

机构地区：[1]北京出入境检验检疫局,北京100026 [2]沈阳农业大学

出　　处：《植物检疫》2011年第3期1-6,共6页Plant Quarantine

基　　金：国家转基因重大专项课题(2008ZX08012-001)

摘　　要：为研制一种可同时检测3种转基因成分BTCry1Ac蛋白、植酸酶蛋白、BTCry1Ah蛋白的蛋白芯片。本文将3种转基因成分蛋白质的单克隆抗体点于化学修饰的基片上,对芯片的表面修饰材料、点样缓冲液成分、封闭液种类、标记二抗使用浓度、反应时间进行优化,并对芯片检测的特异性、灵敏度进行测定。结果表明:研制芯片的表面为环氧修饰,使用含0.1%BSA和30%甘油的点样缓冲液和含1%BSA的封闭液,每次反应时间为30min时,有最佳反应结果;其检测灵敏度为:BTCry1Ac蛋白35ng/mL、植酸酶蛋白20ng/mL、BTCry1Ah蛋白30ng/mL,信号稳定;本研究建立的抗体蛋白芯片检测法可同时检测3种转基因蛋白,并具有较高的灵敏性、特异性和可靠性。The protein chip was prepared for detecting the three transgenic proteins： BT Cryl Ac, Phytase and BT Cryl Ah. Three kinds of monoclonal antibodies corresponding to the proteins were dotted on the chemically modi- fied glass slides, superficial treatment process, spotting concentration, blocking solution type, the concentration of labeled secondary antibody and reaction time were optimized. The detection specificity, and sensitivity were verified. Under the following conditions can obtain the optimal resuh：Surface of the chip was modified by epoxy, spotting buffer contained 0.1% BSA and 30% glycerol, the blocking solution contained 1% BSA, each reaction time was 30minute. The detection sensitivity of 3 proteins were Cryl Ac protein 35ng/mL, phytase protein 20ng/ mL, BT Cryl Ah protein 30ng/mL. Signal of duplicate detection was stability. The detection method of antibody protein chip had high sensitivity, specificity, and reliability. The protein chip reported here can be used to detecte 3 kinds of transgenic proteins. It can be used for GMO detection.

关 键 词：蛋白芯片 转基因蛋白 检测 

分 类 号：Q789[生物学—分子生物学]