A comparison of the accuracy of iTRAQ quantification by nLC-ESI MSMS and nLC-MALDI MSMS methods

Sally L. Shirran, Catherine H. Botting

Research output: Contribution to journalArticlepeer-review

48 Citations (Scopus)

Abstract

The accuracy of quantification obtained using the iTRAQ labelling methodology for measuring protein ratios more extreme than 1 1 was investigated A comparison of nLC-ESI MSMS and nLC-MALDI MSMS analysis routes was performed A fixed concentration of a standard six protein mix was spiked with two proteins at a range of concentrations The two data analysis programmes, Mascot and ProteinPlot Paragon, were also compared Whilst the lower ratios could be measured accurately, greater discrepancies were seen for the higher ratios, particularly by nLC-ESI MSMS Filtering out the weaker reporter ion signals improved the accuracy of the ratios this is likely due to several factors which are explored in more detail Overall, analysis by nLC-MALDI MSMS followed by Mascot interpretation gave the most accurate results (C) 2010 Elsevier B V All rights reserved.

Original languageEnglish
Pages (from-to)1391-1403
Number of pages13
JournalJournal of Proteomics
Volume73
Issue number7
DOIs
Publication statusPublished - 7 May 2010

Keywords

  • iTRAQ quantification
  • nLC-ESI MSMS
  • nLC-MALDI MSMS
  • Mascot
  • ProteinPilot
  • CODED AFFINITY TAGS
  • MASS-SPECTROMETRY
  • QUANTITATIVE-ANALYSIS
  • SHOTGUN PROTEOMICS
  • ISOBARIC TAGS
  • PROTEINS
  • ALGORITHM
  • PEPTIDES
  • COMPLEX

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