Details

Autor(en) / Beteiligte

• 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

Titel

Robust, reproducible and quantitative analysis of thousands of proteomes by micro-flow LC-MS/MS

Ist Teil von

• Nature communications, 2020-01, Vol.11 (1), p.157-157, Article 157

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2020

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.