Robust, reproducible and quantitative analysis of thousands of proteomes by micro-flow LC–MS/MS
2020-01-09
Yangyang Bian, Runsheng Zheng, Florian P. Bayer, Cassandra Wong, Yun-Chien Chang, Chen Meng, Daniel P. Zolg, Maria Reinecke, Jana Zecha, Svenja Wiechmann, Stephanie Heinzlmeir, Johannes Scherr, Bernhard Hemmer, Mike Baynham, Anne-Claude Gingras, Oleksandr Boychenko, Bernhard Kuster
Nat Commun., 2020, 11, 157
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 × 150mm 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.
Speaker: Prof. Dr. Thomas Carell
Ludwig-Maximilians-Universität München
Institut für Chemische Epigenetik (ICEM)
Department of Chemistry
Office:
Würmtalstrasse 201
81377 Munich
Germany
Mailing address:
Butenandtstr. 5 - 13
81377 Munich
Germany
Phone: +49 (0)89 2180-77750
Fax: +49 (0)89 2180-77756
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Managing Director: Dr. Oliver Baron
Institute for Chemical Epigenetics Munich (ICEM)
Ludwig-Maximilians-Universität München
Butenandtstr. 5 - 13
81377 Munich Germany
Phone: +49 (0)89 2180-77661
Fax: +49 (0)89 2180-77651
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Secretary: Birgit Carell
Institute for Chemical Epigenetics Munich (ICEM)
Office:
Würmtalstrasse 201, Haus L - EG / Room: 007
81377 Munich
Germany
Mailing address:
Butenandtstr. 5 - 13
81377 Munich
Germany
Phone: +49 (0)89 2180-77751
Fax: +49 (0)89 2180-77881
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Prof. Dr. Hendrik Zipse (Spokesman)
LMU Munich
Department of Chemistry
Butenandtstr. 5 - 13
House F, Room 3.084
81377 Munich, Germany
Phone: +49 89 2180 77737
Fax: +49 89 2180-77738
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Dr. Stefanie Kellner (Deputy Spokeswoman)
LMU Munich
Department of Chemistry
Butenandtstr. 5 - 13
House F, Room 1.054 und House B, Room 2.073
81377 Munich Germany
Phone: +49 89 2180 77737
This email address is being protected from spambots. You need JavaScript enabled to view it.
Dr. Martin Sumser (Coordinator)
LMU Munich
Department of Chemistry
Butenandtstr. 5 - 13
House F, Room 4.086
81377 Munich Germany
Phone: +49 89 2180 77765
This email address is being protected from spambots. You need JavaScript enabled to view it.
Contact IRTG 1309 related
Email: This email address is being protected from spambots. You need JavaScript enabled to view it.