Proteomics

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High-flow chromatography for reproducible and high-throughput quantification of proteomes


ABSTRACT: This study evaluates liquid-chromatography operated at 50 ul/min using 1 mm diameter columns coupled online to tandem mass spectrometry (LC-MS/MS) for proteome analysis. Data from >1,000 human cell line, tissue, body fluid, affinity- and phospho-proteomes demonstrate excellent chromatographic (<0.3 % CV) and quantitative (<7.5% CV) reproducibility. Combined with sample multiplexing by tandem mass tags, the system allows profiling of 11 proteomes in 16 hours to a depth of 8,000 proteins.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive HF-X, Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Urine, Placenta, Blood Plasma, Cerebrospinal Fluid, Epithelial Cell, Cell Culture, Hela Cell

SUBMITTER: Yangyang Bian  

LAB HEAD: Bernhard Kuster

PROVIDER: PXD015087 | Pride | 2019-12-09

REPOSITORIES: Pride

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Publications


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 analysi  ...[more]

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