NativeMS

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. Collision-induced dissociation can also be performed before or after the IM cell, even with SID installed. This instrument is mainly used for studies of large protein complexes. In addition, we have a linear field ion mobility cell that can be installed and used to make direct collision cross section measurements without the need for calibration, as is necessary for TWIMS measurements.

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 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. Collision-induced dissociation can also be performed before or after the IM cell, even with SID installed. 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 studies of large protein complexes. This instrument also has a linear field ion mobility cell that can be installed and used to make direct collision cross section measurements without the need for calibration.

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.

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.

A 15 T Fourier Transform Ion Cyclotron Resonance (FT-ICR) manufactured by Bruker. This instrument has a trapped ion mobility (TIMS) funnel at the front of the instrument to allow for high-resolution ion mobility (R~200). An SID device for this instrument has been designed and can be installed in place of the collision cell for analysis of large proteins and protein complexes. The ultra-high mass accuracy and ultra-high resolution make this instrument ideal for studying complex systems. A high-mass quadrupole is being designed for this instrument to allow selection up to 32 000 m/z. This instrument is a shared resource housed within the Campus Chemical Instrument Center. This instrument was supported by NIH Award Number Grant S10 OD018507.

The Waters Select Series Cyclic IMS instrument has high sensitivity, high m/z resolution (>100k FWHM) and a unique cyclic ion mobility (cIM) allowing increased ion mobility resolution via multiple ion mobility passes. The instrument will be modified to include surface-induced dissociation, which will be tightly integrated with the system via collaboration with the manufacturer. This instrument was purchased in part with supplement funds from P41GM128577 for method development for characterization of Alzheimer's Disease related aggregates and oligomers.

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.

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.

Mass photometry measures mass at the single molecule level, providing insights into the composition and function of even complex samples and molecular mechanisms. This unique technology can be used to detect and characterise proteins, nucleic acids, lipids and / or sugars, providing information on structure (oligomeric state, modification), homogeneity, and function (quantifying interactions) – all in a matter of minutes, and using tiny amounts of sample. The benchtop Refeyn mass photometer complements existing structural biology equipment at Ohio State by offering interference spectrometry measurements of protein complexes. The mass photometry equipment is housed in the Wysocki lab for use in the P41 program by its Driving Biomedical Projects and Collaboration and Service projects. 

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.