ProteoChip: A highly sensitive protein microarray prepared by a novel method of protein, immobilization for application of protein-protein interaction studies
ProteoChip: A highly sensitive protein microarray prepared by a novel method of protein, immobilization for application of protein-protein interaction studies
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DOI：
10.1002/pmic.200300541
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发表时间：
2003-12-01
期刊：
影响因子：
3.4
通讯作者：
Han, MH
中科院分区：
文献类型：
--
作者：
Lee, Y;Lee, EK;Han, MH
我们开发了一种高灵敏度的微阵列蛋白质芯片ProteoChip，该芯片涂有ProLinker(TM)，这是一种具有双功能偶联特性的新型杯冠衍生物，可以有效地将捕获的蛋白质固定在固体基质上，并使蛋白质相互作用的高通量分析成为可能。石英晶体微天平分析表明，单克隆抗体（mAb）和抗原（Ag）结合在ProLinker(TM) B包覆的传感器表面金膜上，可用于Ab-Ag相互作用的研究。ProteoChip是一种涂有ProLinker(TM) A的胺化玻片，也被证明可用于通过微阵列仪制备高密度阵列点，以及通过直接或夹心荧光免疫分析法分析分析物Ags。ProteoChip的检测灵敏度低至1-10飞图/mL分析物蛋白，可用于肿瘤标志物的检测。ProteoChip也可用于研究直接的蛋白质相互作用，如整合素-细胞外基质蛋白质相互作用的分析所证明的那样。这些实验结果表明，ProteoChip是开发基于芯片的先导筛选微阵列的强大工具，可用于监测蛋白质-蛋白质相互作用（即药物靶点）以及需要高检测灵敏度的生物标志物分析。
We have developed a highly sensitive microarray protein chip, ProteoChip, coated with ProLinker(TM), novel calixcrown derivatives with a bifunctional coupling property that permits efficient immobilization of capture proteins on solid matrixes and makes high-throughput analysis of protein-protein interactions possible. The analysis of quartz crystal microbalance showed that both monoclonal antibody (mAb) and antigen (Ag) bound to the gold film of the sensor surface coated with ProLinker(TM) B and that it is useful for studies of Ab-Ag interactions. ProteoChip, aminated glass slide coated with ProLinker(TM) A, was also demonstrated to be useful for preparation of high-density array spots by using a microarrayer and for analysis of analyte Ags either by direct or sandwich methods of fluorescence immunoassay. The detection sensitivity of ProteoChip was as low as 1-10 femtogram/mL of analyte protein, useful for detection of tumor markers. ProteoChip was also useful for studies of direct protein-protein interactions as demonstrated by analysis of integrin-extracellular matrix protein interaction. These experimental results suggest that ProteoChip is a powerful tool for development of chip-based lead screening microarrays to monitor protein-protein interactions (i.e. drug target) as well as for biomarker assays which require high detection sensitivity.