A review on mass spectrometry-based quantitative proteomics: Targeted and data independent acquisition

Anal Chim Acta. 2017 Apr 29:964:7-23. doi: 10.1016/j.aca.2017.01.059. Epub 2017 Feb 2.


Mass spectrometry (MS) based proteomics have achieved a near-complete proteome coverage in humans and in several other organisms, producing a wealth of information stored in databases and bioinformatics resources. Recent implementation of selected/multiple reaction monitoring (SRM/MRM) technology in targeted proteomics introduced the possibility of quantitatively follow-up specific protein targets in a hypothesis-driven experiment. In contrast to immunoaffinity-based workflows typically used in biological and clinical research for protein quantification, SRM/MRM is characterized by high selectivity, large capacity for multiplexing (approx. 200 proteins per analysis) and rapid, cost-effective transition from assay development to deployment. The concept of SRM/MRM utilizes triple quadrupole (QqQ) mass analyzer to provide inherent reproducibility, unparalleled sensitivity and selectivity to efficiently differentiate isoforms, post-translational modifications and mutated forms of proteins. SRM-like targeted acquisitions such as parallel reaction monitoring (PRM) are pioneered on high resolution/accurate mass (HR/AM) platforms based on the quadrupole-orbitrap (Q-orbitrap) mass spectrometer. The expansion of HR/AM also caused development in data independent acquisition (DIA). This review presents a step-by-step tutorial on development of SRM/MRM protein assay intended for researchers without prior experience in proteomics. We discus practical aspects of SRM-based quantitative proteomics workflow, summarize milestones in basic biological and medical research as well as recent trends and emerging techniques.

Keywords: Data independent acquisition (DIA); Parallel reaction monitoring (PRM); Selected/multiple reaction monitoring (SRM/MRM); Sequential windowed acquisition of all theoretical fragmentation spectra (SWATH); Tandem mass spectrometry (MS/MS); Targeted quantitative proteomics tutorial.

Publication types

  • Review

MeSH terms

  • Humans
  • Mass Spectrometry*
  • Protein Processing, Post-Translational
  • Proteome
  • Proteomics / methods*
  • Reproducibility of Results


  • Proteome