期刊
NATURE METHODS
卷 14, 期 9, 页码 921-+出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/NMETH.4398
关键词
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资金
- Institut Merieux
- US National Institutes of Health (NIH) [R01GM094231]
- Swiss National Science Foundation (SNSF) [P2EZP3 162268, 31003A_166435]
- SNSF Ambizione grant [PZ00P3_161435]
- ERC [AdG-233226, AdG-670821]
- PhosphonetX project of SystemsX.ch
- Swiss National Science Foundation (SNF) [PZ00P3_161435, P2EZP3_162268] Funding Source: Swiss National Science Foundation (SNF)
Liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) is the main method for high-throughput identification and quantification of peptides and inferred proteins. Within this field, data-independent acquisition (DIA) combined with peptide-centric scoring, as exemplified by the technique SWATH-MS, has emerged as a scalable method to achieve deep and consistent proteome coverage across large-scale data sets. We demonstrate that statistical concepts developed for discovery proteomics based on spectrum-centric scoring can be adapted to large-scale DIA experiments that have been analyzed with peptide-centric scoring strategies, and we provide guidance on their application. We show that optimal tradeoffs between sensitivity and specificity require careful considerations of the relationship between proteins in the samples and proteins represented in the spectral library. We propose the application of a global analyte constraint to prevent the accumulation of false positives across large-scale data sets. Furthermore, to increase the quality and reproducibility of published proteomic results, well-established confidence criteria should be reported for the detected peptide queries, peptides and inferred proteins.
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