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Cyclic sample pooling using two‐dimensional liquid chromatography system enhances coverage in shotgun proteomics
Author(s) -
Kawashima Yusuke,
Satoh Mamoru,
Saito Tatsuya,
Matsui Takashi,
Nomura Fumio,
Matsumoto Hiroyuki,
Kodera Yoshio
Publication year - 2013
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.2864
Subject(s) - shotgun proteomics , chemistry , chromatography , shotgun , pooling , proteomics , sample (material) , sample preparation , artificial intelligence , biochemistry , computer science , gene
We report a cyclic sample pooling technique devised in two‐dimensional liquid chromatography–electrospray ionization mass spectrometry (LC‐ESI‐MS) shotgun proteomics that renders deeper proteome coverage; we combined low pH reversed‐phase (RP) LC in trifluoroacetic acid in the first dimension, followed by cyclic sample pooling of the eluate and low‐pH RP‐LC in formic acid in the second dimension. The new protocol has a significantly higher resolving power suitable for LC‐ESI‐MS/MS shotgun proteomics. Copyright © 2013 John Wiley & Sons, Ltd.