Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis

Dingyin Tao, Guijie Zhu, Liangliang Sun, Junfeng Ma, Zhen Liang, Weibing Zhang, Lihua Zhang, Yukui Zhang

Research output: Contribution to journalArticle

Abstract

Microcolumn RPLC (μRPLC) is one of the optimum separation modes for shotgun proteomic analysis. To identify as many proteins as possible by MS/MS, the improvement on separation efficiency and peak capacity of μRPLC is indispensable. Although the increase in column length is one of the effective solutions, the preparation of a long microcolumn is rather difficult due to the high backpressure generated during the packing procedure. In our recent work, through connecting microcolumns of 5, 10, and 15 cm length via unions with minimal dead volume, long microcolumns with length up to 30 cm were obtained, with which 318 proteins were identified from proteins extracted from Escherichia coli by μRPLC-ESI MS/MS, and similar distributions of Mw and pI were found with single and various coupled microcolumns. Furthermore, by using MS/MS with improved sensitivity, with such a serially coupled 30 cm long microcolumn, 1692 proteins were identified within 7 h from rat brain tissue, with false positive rate (FPR)

Original languageEnglish (US)
Pages (from-to)2029-2036
Number of pages8
JournalProteomics
Volume9
Issue number7
DOIs
StatePublished - Apr 2009
Externally publishedYes

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Liquid chromatography
Firearms
Reverse-Phase Chromatography
Proteomics
Proteins
Escherichia coli Proteins
Escherichia coli
Rats
Brain
Tissue

Keywords

  • μRPLC MS/MS
  • Serially coupled microcolumn
  • Shotgun proteome analysis

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

Cite this

Tao, D., Zhu, G., Sun, L., Ma, J., Liang, Z., Zhang, W., ... Zhang, Y. (2009). Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis. Proteomics, 9(7), 2029-2036. https://doi.org/10.1002/pmic.200800649

Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis. / Tao, Dingyin; Zhu, Guijie; Sun, Liangliang; Ma, Junfeng; Liang, Zhen; Zhang, Weibing; Zhang, Lihua; Zhang, Yukui.

In: Proteomics, Vol. 9, No. 7, 04.2009, p. 2029-2036.

Research output: Contribution to journalArticle

Tao, D, Zhu, G, Sun, L, Ma, J, Liang, Z, Zhang, W, Zhang, L & Zhang, Y 2009, 'Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis', Proteomics, vol. 9, no. 7, pp. 2029-2036. https://doi.org/10.1002/pmic.200800649
Tao, Dingyin ; Zhu, Guijie ; Sun, Liangliang ; Ma, Junfeng ; Liang, Zhen ; Zhang, Weibing ; Zhang, Lihua ; Zhang, Yukui. / Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis. In: Proteomics. 2009 ; Vol. 9, No. 7. pp. 2029-2036.
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