Profiling of cytokine expression by biotin-labeled-based protein-arrays
Profiling of cytokine expression by biotin-labeled-based protein-arrays
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DOI：
10.1002/pmic.200300530
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发表时间：
2003-09-01
期刊：
影响因子：
3.4
通讯作者：
Huang, RP
中科院分区：
文献类型：
--
作者：
Lin, Y;Huang, RC;Huang, RP
蛋白质表达的全球分析在基础研究和患者护理中具有很大的前景。以前，我们证明，多种细胞因子可以同时检测使用酶联免疫吸附测定蛋白质阵列系统具有高灵敏度和特异性。在本文中，我们描述了一种基于生物素标记的蛋白质阵列系统，同时检测生物样品中的多种细胞因子。在这种新方法中，来自各种生物来源的蛋白质被生物素标记。然后将生物素标记的蛋白质与抗体芯片一起孵育。目标蛋白质被点在抗体芯片上的阵列抗体捕获。使用Cy 3-或Cy 5-缀合的链霉亲和素检测靶蛋白的存在，并通过激光扫描仪对信号成像。该系统还可以很容易地适应于双色结合测定，允许在同一芯片上相对于参考样品测量测试样品中的蛋白质水平。为了证明其潜在的应用，我们应用该技术来分析雌激素受体（ER）+和ER-细胞中的人细胞因子、趋化因子、生长因子、血管生成因子和蛋白酶。这些结果表明，基于生物素标记的抗体芯片技术可以为研究和临床目的提供分析数百或数千种蛋白质的实用和强大的手段。
Global analysis of protein expression holds great promise in basic research and patient care. Previously we demonstrated that multiple cytokines could be detected simultaneously using an enzyme-linked immunosorbent assay protein array system with high sensitivity and specificity. In this paper, we described a biotin-labeled-based protein array system to detect multiple cytokines simultaneously from biological samples. In this new approach, proteins from A variety of biological sources are labeled with biotin. The biotin-labeled proteins are then incubated with antibody chips. Targeted proteins are captured by the array antibodies spotted on the antibody chips. The presence of targeted proteins is detected using Cy3- or Cy5-conjugated streptavidin and signals are imaged by laser scanner. The system also can be easily adapted to a two-color binding assay, allowing measurement of the levels of proteins in a test sample, with respect to a reference sample at the same chip. To demonstrate its potential applications, we applied this technology to profile human cytokines, chemokines, growth factors, angiogenic factors and proteases in estrogen receptor (ER)+ and ER- cells. These results suggest that biotin-labeled-based antibody chip technology can provide a practical and powerful means of profiling hundreds or thousands of proteins for research and clinical purposes.