Absolute multiplexed protein quantification using QconCAT technology.
Absolute multiplexed protein quantification using QconCAT technology.
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DOI：
10.1007/978-1-61779-885-6_18
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发表时间：
2012-01-01
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影响因子：
--
通讯作者：
Beynon, Robert J
中科院分区：
文献类型：
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作者：
Brownridge, Philip J;Harman, Victoria M;Beynon, Robert J
除了蛋白质鉴定外，蛋白质定量正成为蛋白质组学实验的重要成果。虽然相对定量技术更普遍，是发现蛋白质组学的核心，但大多数分析需要绝对定量。系统生物学的发展也增加了对输入模型的绝对蛋白质丰度值的需求。qconcat是通过将从目标蛋白中提取的肽序列连接到人工蛋白中而产生的。QconCAT作为内部标准的来源，可以对多种蛋白质进行平行绝对定量。qconcat通常应用于靶向蛋白质组学工作流程，因此受益于这些方法的更高灵敏度和更宽的动态范围。在本章中，我们讨论了QconCAT的设计、构造、表达和部署，以及多重绝对量化所需的实验结果。
In addition to protein identification, protein quantification is becoming a key output of proteomic experiments. Although relative quantification techniques are more commonplace and central to discovery proteomics, most assays require absolute quantification. The growth in systems biology has also increased the demand for absolute protein abundance values for input into models. QconCATs are created by concatenating peptide sequences taken from the target proteins into artificial proteins. The QconCAT acts as a source of internal standards and enables parallel absolute quantification of multiple proteins. QconCATs are typically applied in targeted proteomic workflows and so benefit from the greater sensitivity and wider dynamic range of these approaches. In this chapter, we discuss the design, construction, expression, and deployment of a QconCAT and the resulting experiments required for multiplex absolute quantification.