Enabling electron-induced fragmentation in tandem mass spectrometry

The primary market focus for the high end mass spectrometer industry is to fully characterize proteins and their post translational modificationsPTMswithin the biopharmaceutical industryThese analyses remain challenging despite major advances in the speedresolution and mass accuracy of modern mass spectrometersA key weakness with current instrumentation for protein characterization lies in the methods used to induce fragmentationThe reliance in particular on collision induced dissociationCIDhas limited such analyses to bottom up workflows of trypsin digested peptides ofresiduesWhen subjected to CIDmany fragile PTMs on these short peptides are lost in complex waysAn alternative fragmentation methodology called electron capture dissociationECDis well known for producing exceptionally clean spectra of entire proteins while also preserving PTMsHoweverthis technology has been feasible only in expensive FTICR mass spectrometersThe difficulty arises from confining enough low energy electrons to efficiently fragment peptide bondswhich has limited the application of ECD in other instrumentsThe e MSion team has developed an efficient ECD technology to confine electrons with a carefully designed magnetic field that operates without affecting the ion flightpath in mass spectrometersOne major advantage of our technology over competing fragmentation techniques such as ETD is speedWe established Phase I feasibility by showing that our ECD technology is fast enough to be used in quadrupole Time of FlightQToFmass spectrometers at speeds compatible with UPLC and ion mobility based separations of complex samplesOur technology also efficiently supports sequencing of proteins as large askDa in seconds while leaving even the most fragile PTMs intactThe proposed Phase II SBIR project will complete the optimization integration of our ECD into Q ToFandapos s to make the operation seamless for two major manufacturers of Q ToFsThe primary commercial goal is to become a value added reseller for upgrading QTofs in Phase IIITo accomplish thisour first Aim is to refine the engineeringsoftware integration and application to middleand top down protein characterizationAimis to work with early adopters in both Biopharma and in proteomics fields to demonstrate the capabilities of the technologyThe third Aim is to further modify the design of the ECD cell to perform Electron Induced DissociationEIDmore efficiently for the characterization of singly charged peptides and glycoproteinsThis entails subtle modifications to the current ECD cell that allows larger quantities of higher energy electrons to flow through the systemCompletion of Aimwill open the market for triple quad mass spectrometerswhich is five times larger than the more expensive Q ToFsThe adoption of our technology will accelerate the ability of many NIH investigators as well as BioPharma to probe disease mechanisms by characterizing macromolecules in complex biological samples with increased accuracy and speedwhile reducing false discoveries Project NarrativeEven with all of the scientific progress we have made to datethe complexity of disease affected tissues still challenges our ability to probe what makes people sickThe goal of this Phase II SBIR project is to extend our Phase I progress toward developing and commercializing a powerful tool for more effectively cutting large biological molecules into identifiable piecesPhase II success will allow us to engagePhase IIIcommercialization partners and customers with a next generation technology that will improve the diagnosis and treatment of diseases ranging from arthritiscancer and diabetes to heart disease and neurodegeneration