Selected reaction monitoring for quantitative proteomics: a tutorial.
Selected reaction monitoring for quantitative proteomics: a tutorial.
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DOI：
10.1038/msb.2008.61
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发表时间：
2008
影响因子：
9.9
通讯作者：
Aebersold, Ruedi
中科院分区：
文献类型：
--
作者：
Lange, Vinzenz;Picotti, Paola;Domon, Bruno;Aebersold, Ruedi
系统生物学依赖于数据集，其中同一组蛋白质在多个样品中被一致地识别和精确地定量，这一要求仅部分地通过当前的蛋白质组学方法实现。选择反应监测（SRM）-也称为多反应监测-是新兴的技术，理想地补充了发现能力的鸟枪战略，其独特的潜力，可靠的定量分析物的复杂混合物中的低丰度。在SRM实验中，通过三重四极杆仪器的两个质量过滤器选择预定义的前体离子及其碎片之一，并随时间进行监测以进行精确定量。一系列跃迁（前体/碎片离子对）与靶向肽的保留时间结合可构成确定性测定。通常，在单个LC-MS实验期间定量大量肽。本教程解释了SRM在定量蛋白质组学中的应用，包括蛋白质型肽的选择以及转换的优化和验证。此外，归一化和各种因素影响的灵敏度和准确度进行了讨论。
Systems biology relies on data sets in which the same group of proteins is consistently identified and precisely quantified across multiple samples, a requirement that is only partially achieved by current proteomics approaches. Selected reaction monitoring (SRM)—also called multiple reaction monitoring—is emerging as a technology that ideally complements the discovery capabilities of shotgun strategies by its unique potential for reliable quantification of analytes of low abundance in complex mixtures. In an SRM experiment, a predefined precursor ion and one of its fragments are selected by the two mass filters of a triple quadrupole instrument and monitored over time for precise quantification. A series of transitions (precursor/fragment ion pairs) in combination with the retention time of the targeted peptide can constitute a definitive assay. Typically, a large number of peptides are quantified during a single LC-MS experiment. This tutorial explains the application of SRM for quantitative proteomics, including the selection of proteotypic peptides and the optimization and validation of transitions. Furthermore, normalization and various factors affecting sensitivity and accuracy are discussed.
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