双荧光噬菌体蛋白芯片检测系统的研究  

Studies on the detection system of dual fluorescence bacteriophage protein microarray

作　　者：赵美玲[1] 钟理[1] 吴策[1] 王海霞[1] 葛昆[1] 张波[1] 

机构地区：[1]河北大学生命科学学院,河北保定071002

出　　处：《河北农业大学学报》2007年第3期62-65,共4页Journal of Hebei Agricultural University

摘　　要：对双荧光噬菌体蛋白芯片技术的检测性能进行验证,旨在获得一种新的、灵敏度高的蛋白质检测手段。将提纯后的展示GAGE7蛋白(肺癌肿瘤标志物)的T7噬菌体蛋白点到芯片上,将1份肺癌病人血清稀释到1∶100,1∶500,1∶2000,1∶5000,1∶10000,1∶50000,分别用6张相同的蛋白芯片进行筛查,用激光共聚焦扫描仪检测芯片上的荧光信号,分析Cy5/Cy3信号强度和血清稀释度的相关性。同时应用ci-ELISA技术在相同试验条件下进行对比试验。结果表明,Cy5/Cy3信号均与血清稀释度呈良好线性关系,与ELISA法所得结果相符,并且在微量检测能力上要优于ELISA技术。因此,双荧光噬菌体蛋白芯片技术可以对生物样品中某微量蛋白的含量进行准确检测,具有高通量、微型化的特点,是检测生物样品中蛋白质的理想工具。A novel high sensitive protein microarray detection system using dual-fluorescent detection method were developed on phage proteins. Purified T7 phage-expressed GAGE7 （a lung cnacer associated protein） proteins were spotted onto six identical microarray chips. Each chip was then detected simultaneously by a series dilution of lung cancer patient serum from 1 ： 100, 1 ： 500, 1 ： 2000, 1 ： 5000, 1 ： 10000 and 1 ： 50000. The chip images were scanned using a microarray scanner and Cy5/Cy3 signals were analyzed for the correlation with the serum dilutions. Meanwhile, a parallel experiment was done using ci-ELISA detection system for the comparison. Our results showed that the protein chips are more sensitive than the ci-ELISA in detection of antibody concentrations, while both systems showed a good liner relationship between the reactive signal and serum dilutions. Therefore, this novel protein chip detection system cna detect the concentration of the micro-proteins in the biological samples accurately. It also has the caracteristics of high-throughput and micromation, which made it the perfect tool for detecting the proteins in biological samples.

关 键 词：双荧光 噬菌体 蛋白芯片 ELISA 

分 类 号：Q81[生物学—生物工程]