Oral Presentation 10th Australian Peptide Conference 2013

Improved accuracy and sensitivity of large-scale labelled quantitation via multi-notch MS3 approach on hybrid quadrupole-Orbitrap-linear ion trap mass spectrometer (#13)

Lin Tang Goh 1
  1. Thermo Scientific, To be confirmed, tbc, Australia

The utility of multiplexed quantitation using isobaric tags (ITRAQ or TMT) has been well documented in the literature. Today such protein quantitation approach has become a first choice method in many proteomics laboratories due to its proven protocol and productivity to facilitate comparative study of up to 10 experiments. On a high resolution mass spectrometer, the quantitative data are obtained from the cyclic targeted MSMS process (data-dependent acquisition) which releases the associated reporter tags for the detected peptides. In most complex sample situation, isolations of target peptides during DDA have been challenged with co-isolation of other peptides within a 2-Da or 3-Da selection window. This invariably leads to unresolvable error in quatitation. A novel approach based on the new hybrid Orbitrap platform (Orbitrap Fusion) offers significant improvement in accuracy and sensitivity in complex labelled quantitation scenarios. This method involves a triple-stage HRMS measurements (MS3) using ion-trap based CID followed by HCD to generate the corresponding reporter tag ions.