Login
- Titel:
Robust, reproducible and quantitative analysis of thousands of proteomes by micro-flow LC-MS/MS.
- Dokumenttyp:
- Article; Journal Article; Research Support, Non-U.S. Gov't
- Autor(en):
- Bian, Yangyang; Zheng, Runsheng; Bayer, Florian P; Wong, Cassandra; Chang, Yun-Chien; Meng, Chen; Zolg, Daniel P; Reinecke, Maria; Zecha, Jana; Wiechmann, Svenja; Heinzlmeir, Stephanie; Scherr, Johannes; Hemmer, Bernhard; Baynham, Mike; Gingras, Anne-Claude; Boychenko, Oleksandr; Kuster, Bernhard
- Abstract:
- 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.
- Zeitschriftentitel:
- Nat Commun
- Jahr:
- 2020
- Band / Volume:
- 11
- Heft / Issue:
- 1
- Volltext / DOI:
- doi:10.1038/s41467-019-13973-x
- PubMed:
- http://view.ncbi.nlm.nih.gov/pubmed/31919466
- Print-ISSN:
- 2041-1723
- TUM Einrichtung:
- Neurologische Klinik und Poliklinik; Poliklinik für Präventive und Rehabilitative Sportmedizin
BibTeX