基于IgG检测为模型的玻璃介质定量蛋白质芯片的研制及评价  

Development and Evaluation of Glass Protein Microarray for Quantitative Detection of Human Serum IgG

作　　者：费嘉[1,2] 马文丽[1] 吴清华[1] 郑文岭[3] 

机构地区：[1]南方医科大学基因工程研究所,广东省生物芯片重点实验室,中国广东广州510515 [2]暨南大学医学院生物化学与分子生物学教研室,中国广东广州510800 [3]华南基因组研究中心

出　　处：《生命科学研究》2008年第2期154-157,共4页Life Science Research

基　　金：广东省科技计划重点引导项目(2005B33101005);广州市花都区科技计划项目(05-HDGY-001)

摘　　要：为研制高通量定量检测的蛋白质芯片,以人血清IgG为研究模型,选择戊二醛基修饰的玻片为载体,蛋白质样品溶解于20%甘油的PBS,由机械手将浓度为0.5g/L人IgG单克隆抗体点样在玻片上,人血清白蛋白为阴性对照,以1%的BSA为封闭液对蛋白质芯片进行封闭,并经相应处理,构建用于定量检测人血清IgG蛋白质芯片.先以不同浓度IgG标准品为待测样品,建立蛋白质芯片方法和ELISA方法的标准曲线,两条标准曲线的R2值分别为0.996和0.994(P<0.01).两种方法的检测人IgG结果有很好的相关性(R2=0.9937,P<0.01),其敏感性均在15.625μg/L.用两种方法分别检测10例人血清IgG,结果显示两种方法的检测的IgG浓度有很好的一致性,以玻片为载体的蛋白质芯片可用于微量生物样品的定量检测.To fabricate quantitative glass microarray antibody-based glass mieroarray has been developed on the principle similar to the traditional ELISA, an and evaluated on the model of human serum IgG. The glass was silanized and modified with glutarardehyde. Goat-anti-human IgG monoclonal antibody was dissolved in PBS with 20% glycerol, and spotted on the glass slides by robot. Human serum albumin （HSA） was as negative control. Then the glass slides were quenched by PBS with 1% bovine serum albumin （BSA）. Thus, fabrication of protein microarray was completed. Two calibration curves were assembled by detection of human IgG standard samples using protein microarray and traditional ELISA. R^2 values for IgG calibration curves were 0.996 and 0.994, respectively （P 〈 0.01 ）. Detection limit of human IgG standard samples is down to 15.625 μg/L, as sensitive as standard ELISA methods. 10 human serum samples were quantified using protein microarray assay and traditional ELISA method. The results showed IgG concentrations in 10 human serum samples were consistant between those from two methods. Our data indicated that the glass protein microarray is a feasible option for quantification in complex biological samples.

关 键 词：蛋白质芯片 免疫球蛋白G 酶联免疫吸附试验 醛基修饰 

分 类 号：Q819[生物学—生物工程]