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Automated Selected Reaction Monitoring Software for Accurate Label-Free Protein Quantification.

Author:
  • Johan Teleman
  • Christofer Karlsson
  • Sofia Waldemarson
  • Karin M Hansson
  • Peter James
  • Johan Malmström
  • Fredrik Levander
Publishing year: 2012
Language: English
Pages: 3766-3773
Publication/Series: Journal of Proteome Research
Volume: 11
Issue: 7
Document type: Journal article
Publisher: The American Chemical Society

Abstract english

Selected reaction monitoring (SRM) is a mass spectrometry method with documented ability to quantify proteins accurately and reproducibly using labeled reference peptides. However, the use of labeled reference peptides becomes impractical if large numbers of peptides are targeted and when high flexibility is desired when selecting peptides. We have developed a label-free quantitative SRM workflow that relies on a new automated algorithm, Anubis, for accurate peak detection. Anubis efficiently removes interfering signals from contaminating peptides to estimate the true signal of the targeted peptides. We evaluated the algorithm on a published multisite data set and achieved results in line with manual data analysis. In complex peptide mixtures from whole proteome digests of Streptococcus pyogenes we achieved a technical variability across the entire proteome abundance range of 6.5-19.2%, which was considerably below the total variation across biological samples. Our results show that the label-free SRM workflow with automated data analysis is feasible for large-scale biological studies, opening up new possibilities for quantitative proteomics and systems biology.

Keywords

  • Immunology in the medical area

Other

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

Professor

Department of Immunotechnology

+46 46 222 14 96

+46 70 247 79 60

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