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

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

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6 Citations (SciVal)

Abstract

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.
Original languageEnglish
Pages (from-to)394-399
JournalJournal of Proteome Research
Volume4
Issue number2
DOIs
Publication statusPublished - 2005

Subject classification (UKÄ)

  • 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

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