Mass spectrometry-based proteomics using Q Exactive, a high-performance benchtop quadrupole Orbitrap mass spectrometer
Mol Cell Proteomics. 2011 Jun 3.
Michalski A, Damoc E, Hauschild JP, Lange O, Wieghaus A, Makarov A, Nagaraj N, Cox J, Mann M, Horning S.
Mass spectrometry-based proteomics has greatly benefitted from enormous advances in highresolution instrumentation in recent years. In particular, the combination of a linear ion trap with the Orbitrap analyzer has proven to be a popular instrument configuration.Complementing this hybrid trap-trap instrument, as well as the standalone Orbitrap analyzer termed Exactive, we here present coupling of a quadrupole mass filter to an Orbitrap analyzer. This Q Exactive instrument features high ion currents due to an S-lens, and fast HCD peptide fragmentation due to parallel filling and detection modes. The image current from the detector is processed by an enhanced Fourier Transformation (eFT) algorithm, doubling mass spectrometric resolution. Together with almost instantaneous isolation and fragmentation, the instrument achieves overall cycle times of 1 s for a top10 HCD method.More than 2,500 proteins can be identified in standard 90 min gradients of tryptic digests of mammalian cell lysates - a significant improvement over previous Orbitrap mass spectrometers. Furthermore, the quadrupole Orbitrap analyzer combination enables multiplexed operation at the MS and MS/MS levels. This is demonstrated in a multiplexed single ion monitoring mode, in which the quadrupole rapidly switches between different narrow mass ranges that are analyzed in a single composite MS spectrum. Similarly, the quadrupole allows fragmentation of different precursor masses in rapid succession, followed by joint analysis of the HCD fragment ions in the Orbitrap analyzer. High performance in a robust benchtop format together with the ability to perform complex multiplexed scan modes make the Q Exactive an exciting new instrument for the proteomics and general analytical communities.