W.M. Keck Foundation Biotechnology Resource Laboratory, School of Medicine, Yale University, New Haven, CT 06510, USA.
Methods. 2013 Jun 15;61(3):287-98. doi: 10.1016/j.ymeth.2013.05.004. Epub 2013 May 21.
Selective or Multiple Reaction monitoring (SRM/MRM) is a liquid-chromatography (LC)/tandem-mass spectrometry (MS/MS) method that enables the quantitation of specific proteins in a sample by analyzing precursor ions and the fragment ions of their selected tryptic peptides. Instrumentation software has advanced to the point that thousands of transitions (pairs of primary and secondary m/z values) can be measured in a triple quadrupole instrument coupled to an LC, by a well-designed scheduling and selection of m/z windows. The design of a good MRM assay relies on the availability of peptide spectra from previous discovery-phase LC-MS/MS studies. The tedious aspect of manually developing and processing MRM assays involving thousands of transitions has spurred to development of software tools to automate this process. Software packages have been developed for project management, assay development, assay validation, data export, peak integration, quality assessment, and biostatistical analysis. No single tool provides a complete end-to-end solution, thus this article reviews the current state and discusses future directions of these software tools in order to enable researchers to combine these tools for a comprehensive targeted proteomics workflow.
选择或多重反应监测(SRM/MRM)是一种液相色谱(LC)/串联质谱(MS/MS)方法,通过分析样品中特定蛋白质的前体离子和其选定的胰蛋白酶肽的碎片离子,实现对其的定量分析。仪器软件已经发展到了一个地步,即通过精心设计的 m/z 窗口的调度和选择,可以在与 LC 耦合的三重四极杆仪器上测量数千个跃迁(一对主要和次要 m/z 值)。一个好的 MRM 测定的设计依赖于以前的 LC-MS/MS 研究中发现阶段的肽谱的可用性。涉及数千个跃迁的手动开发和处理 MRM 测定的繁琐方面促使开发了自动化该过程的软件工具。已经开发了软件包用于项目管理、测定开发、测定验证、数据导出、峰积分、质量评估和生物统计学分析。没有单一的工具提供完整的端到端解决方案,因此本文综述了这些软件工具的当前状态,并讨论了它们的未来发展方向,以便研究人员能够将这些工具结合起来,实现全面的靶向蛋白质组学工作流程。