Fröhlich Thomas, Arnold Georg J
Laboratory for Functional Genome Analysis LAFUGA, Gene Center, Ludwig-Maximilians-University, Munich, Germany.
Methods Mol Biol. 2011;790:141-64. doi: 10.1007/978-1-61779-319-6_11.
Selected reaction monitoring (SRM) is one of the most powerful techniques for the relative and absolute quantification of proteins from complex protein mixtures. In contrast to traditional protein quantification methods such as ELISAs or RIAs, the SRM method uses mass spectrometry for detection. Further benefits of SRM are as follows: (1) high specificity and sensitivity; (2) large linear dynamic range of at least three orders of magnitude; and (3) the possibility to quantify multiple proteins simultaneously in a single MS run from an individual sample. To perform SRM-based protein quantification reliably, a careful design of the assay is essential, and several pitfalls must be avoided. The aim of this chapter is to help SRM newcomers to establish SRM-based protein quantification assays and discuss an overview of typical work flows that are applied during SRM assay development.
选择反应监测（SRM）是对复杂蛋白质混合物中的蛋白质进行相对和绝对定量分析最强大的技术之一。与传统的蛋白质定量方法如酶联免疫吸附测定（ELISA）或放射免疫测定（RIA）不同，SRM方法使用质谱进行检测。SRM的其他优点如下：（1）高特异性和灵敏度；（2）至少三个数量级的大线性动态范围；（3）能够在一次质谱运行中同时对单个样品中的多种蛋白质进行定量。为了可靠地进行基于SRM的蛋白质定量分析，仔细设计分析方法至关重要，并且必须避免一些陷阱。本章的目的是帮助初次接触SRM的人员建立基于SRM的蛋白质定量分析方法，并讨论SRM分析方法开发过程中应用的典型工作流程概述。
