Robust, reproducible and quantitative analysis of thousands of proteomes by micro-flow LC-MS/MS.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
09 01 2020
09 01 2020
Historique:
received:
14
09
2019
accepted:
10
12
2019
entrez:
11
1
2020
pubmed:
11
1
2020
medline:
25
3
2020
Statut:
epublish
Résumé
Nano-flow liquid chromatography tandem mass spectrometry (nano-flow LC-MS/MS) is the mainstay in proteome research because of its excellent sensitivity but often comes at the expense of robustness. Here we show that micro-flow LC-MS/MS using a 1 × 150 mm column shows excellent reproducibility of chromatographic retention time (<0.3% coefficient of variation, CV) and protein quantification (<7.5% CV) using data from >2000 samples of human cell lines, tissues and body fluids. Deep proteome analysis identifies >9000 proteins and >120,000 peptides in 16 h and sample multiplexing using tandem mass tags increases throughput to 11 proteomes in 16 h. The system identifies >30,000 phosphopeptides in 12 h and protein-protein or protein-drug interaction experiments can be analyzed in 20 min per sample. We show that the same column can be used to analyze >7500 samples without apparent loss of performance. This study demonstrates that micro-flow LC-MS/MS is suitable for a broad range of proteomic applications.
Identifiants
pubmed: 31919466
doi: 10.1038/s41467-019-13973-x
pii: 10.1038/s41467-019-13973-x
pmc: PMC6952431
doi:
Substances chimiques
Peptides
0
Proteome
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
157Subventions
Organisme : CIHR
ID : CIHR FDN 143301
Pays : Canada
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