Optimization of LC/MS Intact /Top-Down Protein Analysis on an Orbitrap Fusion Mass Spectrometer

Methods Sample Preparation and Liquid Chromatography Protein standards (Ubiquitin, Carbonic anhydrase and Enolase) were purchased from Sigma Aldrich. Proteins were infused at 1pmol/ul at a flow rate 5ul/min for direct infusion experiments. Recombinant Human TROP2 protein (extracellular domain) was purchased…

Thermo Scientific™ Proteome Discoverer™ using a XlinkX software node. Methods: Different amine-reactive, homobifuctional crosslinkers including disuccinimidyl Results: For both DSSO and BuUrBu, we identified over 40 BSA inter-crosslinked peptides 2 CID for DSSO. We also to less than(DSS), 20 using…

evaluated traditional non-cleavable and MS-cleavable crosslinkers for crosslinked peptide analysis using an Orbitrap Fusion Lumos mass spectrometer. For MS-cleavable crosslinkers, we also compared different types of fragmentation (CID, ETD) and levels of tandem mass spectrometry (MS2 vs. MS3). Our data…

Optimization of RP-LC-MS Top-down Protein Analysis on an Orbitrap Fusion Lumos Tribrid MS with the Advanced Peak Determination Algorithm Vlad Zabrouskov1, Luca Fornelli2, Ryan Fellers2, Kristina Srzentic2, Joshua Silveira1, Helene Cardasis1, Graeme C. McAlister1, Christian Thoeing3, Derek Bailey1, Andreas Kuehn3,…