MicroSPE-nanoLC-ESI-MS/MS using 10-microm-i.d. silica-based monolithic columns for proteomics

Anal Chem. 2007 Jan 15;79(2):540-5. doi: 10.1021/ac061603h.

Abstract

Silica-based monolithic capillary columns (25 cm x 10 microm i.d.) with integrated nanoESI emitters have been developed to provide high-quality and robust microSPE-nanoLC-ESI-MS analyses. The integrated nanoESI emitter adds no dead volume to the LC separation, allowing stable electrospray operation at flow rates of approximately 10 nL/min. In an initial application with a linear ion trap MS, we identified 5510 unique peptides that covered 1443 distinct Shewanella oneidensis proteins from a 300-ng tryptic digest sample in a single 4-h LC-MS/MS analysis. The use of an integrated monolithic ESI emitter provided enhanced resistance to clogging and provided good run-to-run reproducibility.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Chromatography, Liquid / instrumentation*
  • Peptides / analysis
  • Proteomics / instrumentation*
  • Reproducibility of Results
  • Shewanella / chemistry
  • Shewanella / metabolism
  • Silicon Dioxide / chemistry
  • Spectrometry, Mass, Electrospray Ionization / instrumentation*
  • Trypsin / chemistry

Substances

  • Peptides
  • Silicon Dioxide
  • Trypsin