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Wysocki Research Group

Director: Vicki Wysocki

Summary:

Under the direction of Vicki Wysocki, Professor of Chemistry and Biochemistry, Ohio Eminent Scholar, Director of the Campus Chemical Instrument Center, and Director of the NIH-funded Resource for Native Mass Spectrometry Guided Structural Biology, the Wysocki group includes graduate and undergraduate students, postdoctoral researchers and research staff,. Research in the Wysocki group is categorized into four broad areas: (1) development and implementation of surface-induced dissociation onto commercial time-of-flight, Orbitrap and FT-ICR instruments, (2) development and application of native mass spectrometry-guided structural biology approaches, (3) multi-omics approaches to biomarker discovery, disease diagnosis and prognosis using proteomics and metabolomics methods coupled with genomics and transcriptomics, and (4) determination of peptide and other fragment ion structures by IR action spectroscopy.

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Instruments

  • SCIEX (Beckman Coulter) CESI 8000 Plus High-Performance Separation-Electrospray Ionization (ESI) Module ( Electrophoresis system )

    The SCIEX (Beckman Coulter) CESI 8000 Plus High-Performance Separation-Electrospray Ionization (ESI) Module integrates ultra-low flow capillary electrophoresis (CE) with ESI to enable high-resolution separation of minute samples for MS analysis. The CESI 8000 Plus can be used to analyze charged and polar analytes and to characterize peptides, proteoforms, post-translational modifications, and proteins.

  • Thermo Scientific Exactive Plus Extended Mass Range (EMR) Orbitrap mass spectrometer ( Tandem mass spectrometer )

    A benchtop Orbitrap instrument with extended mass range (EMR), which enables analysis of large molecules (m/z 350-20,000) with high mass accuracy and is equipped with a variable pressure higher-energy collision-induced dissociation (HCD) cell. This instrument has been modified to include a high-mass range quadrupole mass filter and a custom surface-induced dissociation (SID) device. The EMR is primarily used for analysis of proteins and protein complexes by both direct infusion and online non-denaturing separation techniques.

  • Thermo Scientific Q Exactive Ultra-High Mass Range (UHMR) Hybrid Quadrupole-Orbitrap mass spectrometer ( Tandem mass spectrometer )

    The Thermo Scientific Q Exactive Ultra-High Mass Range (UHMR) Hybrid Quadrupole-Orbitrap instrument enables analysis of extra-large molecules (m/z 350-80,000) with high mass accuracy. The UHMR is equipped with a higher-energy collision-induced dissociation (HCD) cell capable of acceleration voltages up to 300 V, as well as in-source trapping capabilities for improved declustering of ions. The Wysocki lab has two of these instruments and they are both modified with custom surface-induced dissociation (SID) devices. The UHMRs are primarily used for analysis of proteins and protein complexes by both direct infusion and online non-denaturing separation techniques. One instrument was purchased with funds from P41GM128577 and the other is part of a direct collaboration with Thermo Scientific.

  • Thermo Scientific Vanquish Duo Ultra-High-Performance Liquid Chromatography (UHPLC) system ( Liquid chromatography instrument )

    Thermo Scientific Vanquish Duo Ultra-High-Performance Liquid Chromatography (UHPLC) system enables online separations coupled with mass spectrometry and is equipped with a binary pump and autosampler. The UHPLC system is primarily used for automated chromatography (e.g., size exclusion, ion exchange, immobilized metal affinity, reversed phase, online buffer exchange) online with MS.

  • Thermo Scientific Vanquish Ultra-High-Performance Liquid Chromatography (UHPLC) system ( Liquid chromatography instrument )

    The Thermo Scientific Vanquish Ultra-High-Performance Liquid Chromatography (UHPLC) system enables online separations coupled with mass spectrometry and is equipped with a binary pump and autosampler. The UHPLC system is primarily used for automated chromatography (e.g., size exclusion, ion exchange, immobilized metal affinity, reversed phase, online buffer exchange) online with MS.

  • Thermo Scientific Orbitrap Elite Hybrid Ion Trap-Orbitrap mass spectrometer ( Ion trap mass spectrometer )

    This hybrid instrument consists of a dual linear ion trap, an HCD cell and a high-field orbitrap analyzer. The orbitrap provides high resolution at up to 240,000. The dual linear ion trap allows faster scan rates. For tagged samples, the HCD cell allows lower m/z scan ranges. This instrument is also equipped with ETD which allows for fragmentation of longer peptides and top-down proteomics studies. The instrument is coupled to a 2D Waters nanoACQUITY which allows LC/LC-MS/MS analysis. It is suitable for many -omics studies.

  • Thermo Scientific Velos Pro Dual-Pressure Linear Ion Trap mass spectrometer ( Ion trap mass spectrometer )

    This dual pressure linear ion trap allows fast scan rates. Along with trap HCD and ETD capabilities, this instrument is ideal for routine complex proteomic analysis. The instrument is currently coupled with a 1D Waters nanoACQUITY UPLC system for LC-MS/MS.

  • Waters Select Series Cyclic Ion-Mobility Separation (IMS) mass spectrometer ( Ion trap mass spectrometer )

    Developed through focused collaboration, the SELECT SERIES Cyclic IMS combines novel cyclic ion mobility separation with state-of-the-art high performance, time-of-flight mass spectrometry, enabling leading researchers to unlock the potential in scientific discovery.

  • Waters Synapt "G1" High Definition Mass Spectrometry (HDMS) mass spectrometer ( Tandem mass spectrometer )

    The first generation Waters Q-TWIM-TOF platform. The high sensitivity of this instrument makes it particularly well suited to the study of low concentration samples. This instrument is primarily used for ion mobility studies of low concentration samples. A surface-induced dissociation (SID) device will be designed to replace the trap cell in this instrument.

  • Waters Synapt G2 High Definition Mass Spectrometry (HDMS) mass spectrometer ( Tandem mass spectrometer )

    A quadrupole/travelling wave ion mobility (TWIM)/time-of-flight mass spectrometer. Ion mobility enables gas-phase ions to be separated by their size, shape, and charge. This affords a dimension of separation orthogonal to the standard m/z analysis. The instrument has been modified with a custom surface-induced dissociation (SID) device which can be placed before the IM region (SID-IM), after the IM region (IM-SID), or on either side of the IM cell to allow for SID-IM-SID. In addition, a linear field ion mobility cell can be installed and used to make direct collision cross section measurements without the need for calibration, as is necessary for TWIMS measurements. This instrument is mainly used for studies of large protein complexes.

  • Waters Synapt G2S High Definition Mass Spectrometry (HDMS) mass spectrometer ( Tandem mass spectrometer )

    A newer generation of Waters Synapt G2 instrument with improved “step-wave” design after the ion source for improved sensitivity. Like the G2 this instrument has been modified with a custom surface-induced dissociation (SID) device that can be placed in the same possible configurations. This instrument is equipped with electron transfer dissociation (ETD) and has also been modified to incorporate a 193 nm laser for ultraviolet photo dissociation (UVPD). It is mainly used for protein complexes and more specifically for coupling SID and UVPD to do complex-down and top-down studies.

Resource Collection

Software

  • Proteome Discoverer by Thermo Fisher Scientific ( Software )

    Software for identification and quantification of proteins in complex biological samples for a wide range of proteomic workflows, from protein and peptide identification to PTM analysis to isobaric mass tagging and both SILAC and label-free quantitation.


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Last updated: 2021-03-22T10:32:05.517-04:00

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