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Peter James

Peter James

Professor

Peter James

Reproducibility of tryptic digestion investigated by quantitative fourier transform ion cyclotron resonance mass spectrometry

Author

  • C Hagman
  • M Ramstrom
  • Maria Bentz
  • Peter James
  • P Hakansson
  • J Bergquist

Summary, in English

In this study, the reproducibility of tryptic digestion of complex solutions was investigated using liquid chromatography Fourier transform ion cyclotron resonance (LC FT-ICR) mass spectrometry. Tryptic peptides, from human cerebrospinal fluid, (CSF) were labeled with Quantification-Using-Enhanced-Signal-Tags (QUEST)-markers, or 1-([H-4]nicotinoyloxy)- and 1-([D-4]nicotinoyloxy)-succinimide ester markers. The analysis was performed on abundant proteins with respect-to-intensity ratios and sequence coverage and obtained by comparing differently labeled components from one or different pools. To interpret the dynamics in the proteome, one must be able to estimate the error introduced in each experimental steps. The intra sample variation due to derivatization was approximately 10%. The inter sample variation depending on derivatization and tryptic digestion was not more than approximately 30%. These experimental observations provide a range for the up- and clown-regulations that are possible to study with electrospray ionization LC FT-ICR mass spectrometry.

Department/s

  • Departments at LTH

Publishing year

2005

Language

English

Pages

394-399

Publication/Series

Journal of Proteome Research

Volume

4

Issue

2

Document type

Journal article

Publisher

The American Chemical Society (ACS)

Topic

  • Medical Engineering

Keywords

  • 1-([D-4]nicotinoyloxy)-succinimide ester markers
  • 1-([H-4]nicotinoyloxy)- and
  • quantification
  • QUEST-markers
  • LC FT-ICR mass spectrometry
  • cerebrospinal fluid
  • tryptic reproducibility

Status

Published

ISBN/ISSN/Other

  • ISSN: 1535-3893