Intact protein analysis provides proteoform-specific understanding of biological phenomena that cannot be achieved by analysis of digested peptides. The 21 Tesla FT-ICR mass spectrometer at the National High Magnetic Field Laboratory (NHMFL) offers high mass resolving power, mass accuracy, dynamic range, and scan rate, and achieves unprecedented performance with respect to intact protein sequence analysis. The 21 T is part of the NHMFL FT-ICR User Facility and is available to all qualified users.
This talk will provide performance benchmarks of the 21 T FT-ICR mass spectrometer for characterization of intact proteins and highlight results from ongoing top-down proteomics projects. Recent efforts have focused on enabling new ion manipulation strategies prior to FTMS. For example, we have enabled chimeric ion loading, high resolution SWIFT ion isolation, proton transfer reactions (PTR) and parallel ion parking (PIP). Use of PTR reactions, when coupled with PIP, combine the majority of the initial protein charge state distribution into just a few charge states. This increases S/N, which enables observation of proteoforms that are not observed if PTR/PIP is not employed. The techniques described facilitate rapid identification of intact proteins and extend the mass range available for top-down proteomics applications.
Dr. Lissa Anderson
Lissa Anderson is research faculty and the director of biological applications for the Ion Cyclotron User Facility of the National High Magnetic Field Laboratory (MagLab). She obtained her PhD at the University of Virginia under Professor Donald Hunt and Dr. Jeffrey Shabanowitz and was a postdoctoral associate of the MagLab’s ICR Program.. Her research on top-down proteomics includes development and application of technologies to perform and control ion-ion chemistry inside the mass spectrometer to improve proteoform detection sensitivity and primary structure determination. More at https://bit.ly/3ofw99v