iTRAQ analysis with Paul ion trap-obstacle solved.

ABSTRACT

Mass spectrometers equipped with ion trap analyzers have been significantly improved due to their high performance and wide application area accompanying the low costs of purchase. Despite several advantages, such as reasonable resolution at low cost, high sensitivity, and capability for multistage analysis, ion traps have an important drawback low mass cutoff during tandem mass spectrometry analysis MS(n). Although the low mass cutoff associated with the ion trap does not seriously obstruct peptide identification, it may cause a serious problem in identification of small molecules (posttranslational modifications, e.g., glycan structures) and quantification of peptides with multiplexed isobaric tag reagents. The presented approach offers the possibility to use isobaric tags for relative and absolute quantification labeling (iTRAQ) for quantitative, proteomic analysis using typical, widely available ion trap devices and manufacturer's software. We have performed series of analyses of standard protein labeled with isobaric tags in various concentration ratios to prove quantitative capabilities of this approach.