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Cyclic sample pooling using two‐dimensional liquid chromatography system enhances coverage in shotgun proteomics |
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| Authors: | Yusuke Kawashima Mamoru Satoh Tatsuya Saito Takashi Matsui Fumio Nomura Hiroyuki Matsumoto Yoshio Kodera |
| Affiliation: | 1. Center for Disease Proteomics, School of Science, Kitasato University, , Sagamihara‐shi, Kanagawa, 252‐0373 Japan;2. Laboratory of Biomolecular Dynamics, Department of Physics, School of Science, Kitasato University, , Sagamihara‐shi, Kanagawa, 252‐0373 Japan;3. Clinical Proteomics Research Center, Chiba University Hospital, , Chiba‐shi, Chiba, 260‐8670 Japan;4. Division of Natural Products Chemistry, Department of Medicinal Resources, Institute of Natural Medicine, University of Toyama, , Toyama‐shi, Toyama, 930‐0194 Japan;5. Department of Molecular Diagnosis (F8) Graduate School of Medicine, Chiba University, , Chiba‐shi, Chiba, 260‐8670 Japan;6. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, , Oklahoma City, OK 73104 USA |
| Abstract: | 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. |
| Keywords: | cyclic sample pooling 2D‐LC‐ESI‐MS shotgun proteomics serum proteins |