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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
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

Abstract 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.

Keywords

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

Other

Published
  • ISSN: 1535-3893
Peter James
E-mail: peter.james [at] immun.lth.se

Professor

Department of Immunotechnology

+46 46 222 14 96

+46 70 247 79 60

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