Speaker
Description
Innovations in mass spectrometry (MS) instrumentation continue to emerge, driven primarily by the need to identify and characterize proteins with greater confidence. Simultaneously, fragmentation schemes are an essential performance component of any MS platform, delivering detailed structural and sequence data necessary for precise identifications. Here we report on the latest advances realized on the new timsOmni platform where novel offline as well as online data dependent acquisition (DDA) experimental workflows are applied for the analysis of different classes of analytes. The diverse operating modes described in this study highlight the exceptional versatility and broad applicability of this novel instrument configuration.
A series of hardware developments are reported including (a) a linear stacked-ring RF ion guide providing efficient desolvation, (b) an ion mobility gate for selecting ions separated in the tims device, (c) a new design of segment Q5 in the Omnitrap accommodating higher electron currents without comprising robustness and (d) an improved AC-ejection method to transfer a wide range of m/z ratios from the collision cell to the TOF analyzer. The range of m/z ratios recorded in single TOF spectra extends from <300 Th to >10,000 Th. An offline ESI source is developed and applied for analysis of protein complexes. Deep sequencing of IgG1 monoclonal antibodies fragmented by various MSn modes is reported. Gas phase reduction of intrachain disulfide bonds is demonstrated for intact mAbs and industrial enzymes using electron-based fragmentation while post translational modifications on histones are identified in MSn experiments. Collision activation followed by mobility separation and electron capture dissociation is performed to map the unfolding process of different protein systems. DDA experiments are performed on a protein standard and digested antibody mixtures. Dynamic control of the accumulation period for quad-selected charge states is shown to improve sequence coverage for all the protein systems and subunits examined.
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