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

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Authors

VIDOVÁ Veronika SPÁČIL Zdeněk

Year of publication 2017
Type Article in Periodical
Magazine / Source Analytica Chimica Acta
MU Faculty or unit

Faculty of Science

Citation
Web https://www.sciencedirect.com/science/article/pii/S0003267017301502?via%3Dihub
Doi http://dx.doi.org/10.1016/j.aca.2017.01.059
Keywords Targeted quantitative proteomics tutorial; Selected/multiple reaction monitoring (SRM/MRM); Parallel reaction monitoring (PRM); Tandem mass spectrometry (MS/MS); Data independent acquisition (DIA)
Description 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. SRMlike 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.
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