Sequential Multiplex Analyte Capturing for Phosphoprotein Profiling
Sequential Multiplex Analyte Capturing for Phosphoprotein Profiling
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DOI：
10.1074/mcp.m110.002709
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发表时间：
2010-11-01
影响因子：
7
通讯作者：
Joos, Thomas O.
中科院分区：
文献类型：
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作者：
Poetz, Oliver;Henzler, Tanja;Joos, Thomas O.
基于微阵列的夹心免疫测定可以同时检测数十种蛋白质。然而，它们在定量大量蛋白质中的应用受到免疫测定本身引起的交叉反应性和不相容性的阻碍。顺序多重分析物捕获解决了这些问题，通过重复探测相同的样品与不同的抗体包被，磁悬浮珠阵列。作为一种小型化的免疫测定形式，基于悬浮珠阵列的测定满足环境分析物理论的标准，并且我们的实验表明分析物浓度没有显著变化。通过探测肿瘤细胞系裂解物中7种不同受体酪氨酸激酶的丰度及其磷酸化程度，并通过测量来自同一腔的同一样品中表皮生长因子受体的复杂磷酸化模式，证明了顺序多重分析物捕获的价值。Molecular & Cellular Proteomics 9：2474-2481，2010.
Microarray-based sandwich immunoassays can simultaneously detect dozens of proteins. However, their use in quantifying large numbers of proteins is hampered by cross-reactivity and incompatibilities caused by the immunoassays themselves. Sequential multiplex analyte capturing addresses these problems by repeatedly probing the same sample with different sets of antibodycoated, magnetic suspension bead arrays. As a miniaturized immunoassay format, suspension bead array-based assays fulfill the criteria of the ambient analyte theory, and our experiments reveal that the analyte concentrations are not significantly changed. The value of sequential multiplex analyte capturing was demonstrated by probing tumor cell line lysates for the abundance of seven different receptor tyrosine kinases and their degree of phosphorylation and by measuring the complex phosphorylation pattern of the epidermal growth factor receptor in the same sample from the same cavity. Molecular & Cellular Proteomics 9:2474-2481, 2010.