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PMID: 18854821 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Selected reaction monitoring for quantitative proteomics: a tutorial.

Molecular systems biology ·Vol. 4 ·2008-00-00 ·Pages 222

Lange V, Picotti P, Domon B, Aebersold R

Abstract

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.

MeSH Terms
Proteomics/methods Systems Biology/methods Tandem Mass Spectrometry/methods
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lange Vinzenz
Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland.
Picotti Paola
Domon Bruno
Aebersold Ruedi
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Article Info
Journal
Molecular systems biology
Abbr.
Mol Syst Biol
ISSN
1744-4292
DOI
10.1038/msb.2008.61
Published
2008-00-00
Epub
2008-00-14
Pages
222
Language
English
Region
England
NLM ID
101235389
PMCID
PMC2583086
Subset
IM
Grants
NHLBI NIH HHS · N01HV28179 · United States
NHLBI NIH HHS · N01-HV-28179 · United States
External Links
PubMed source DOI: 10.1038/msb.2008.61 PMC Full Text View journal details
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