【摘要】 目的应用金磁微粒标记蛋白质技术,建立可目视化蛋白质芯片检测体系,比较金磁微粒和胶体金标记蛋白质技术应用于蛋白质芯片检测效果的优劣。方法将人IgG点制于环氧基修饰的玻片上,分别与金磁微粒和胶体金标记的羊抗人IgG温育,银染显色,肉眼观察并用普通扫描仪记录结果。结果基于金磁微粒的蛋白质芯片人IgG最佳点样浓度为0.2mg/ml,37℃温育2h,银染10～15min,检测结果信噪比高;基于胶体金的蛋白质芯片人IgG最佳点样浓度为0.1mg/ml,37℃温育1h,银染15～20min,检测结果信噪比高。结论金磁微粒标记蛋白质技术应用于蛋白质芯片的检测,具有和胶体金一致的可目视化检测效果,且其标记技术简单,标记的蛋白质可定量。更多还原

【Abstract】 Objective To develop a visible detection system prepared by protein microarrays which was based on GoldMag particles-labeled technique and compare the results of detection using the protein which was labeled with GoldMag particles and colloidal gold.Methods Human IgG was printed on the glass slides modified with epoxy groups,and goat anti-human IgG conjugated with GoldMag particles and colloidal gold respectlively was then added on the slide.The glass slides were incubated,and then the black images of microarray spots were produced by immuno-gold-silver staining method and observed by naked eyes and recorded with common flatbed scanner.Results The high signal-to-noise ratio could be obtained when the optimized procedures of GoldMag particles-labeling probe were introduced to the protein chip.The conditions of optimum assay were as follows:the spotting concentration of human IgG was 0.2 mg/ml,the glass slides were incubated at 37 ℃ for 2 hour to immobilize human IgG,and the silver enhancement time was 10 min-15 min.Parallelly,the optimized conditions for colloidal gold were as follows:the spotting concentration of human IgG was 0.1 mg/ml,the slides were incubated at 37 ℃ for 1 hour to immobilize human IgG,and the silver enhancement time was 15 min-20 min.Conclusions GoldMag particles-labeled protein technique is comparable to colloidal gold in applying to protein microarrays.The method is considerably simple and practical,and the protein labeled by GoldMag particles could be quantitative.更多还原