CAREER: Advancing Ion-ion Reactions to Capture the Combinatorial Molecular Features of the Glycocode

Glycoproteins play critical roles in health and disease, from cancer to infection to immune function, but current tools struggle to define heterogenous populations of glycoproteins with molecular precision. This project uses tandem mass spectrometry for developing new methods to characterize complex glycoproteins. The experimental and computational methods should improve upon existing approaches to identify chemical changes in differing glycosylation states, potentially enabling new biological discoveries, contributing to the production of glycoprotein-based pharmaceuticals, and benefiting societal health. As part of this project, the team will expand access to science education by growing the University of Washington Chemistry Frontiers outreach program, which connects students from all backgrounds to exciting topics in chemistry, and by creating openly accessible teaching materials on bioanalytical mass spectrometry with hands-on demonstrations for use by students and educators across the nation. Together, these efforts will advance scientific literacy, broaden participation in Science, Technology, Engineering, and Mathematics (STEM), and help train the next generation of scientists in advanced biotechnology approaches for careers in both academic and industrial research. This proposal aims to improve the ability of mass spectrometry to define the compositional and structural features of complex and heterogenous glycoproteoforms. The project will optimize instrumentation to improve glycopeptide and glycoprotein characterization with combinations of collision-, electron-, and photon-based activation in the gas phase. These advances will be coupled with new informatic strategies to interpret highly dimensional data types and extract useful molecular signatures from complex spectra. These methodologies will aid both in mapping combinatorial modification states of glycoproteins and in differentiating structural features of glycan isomers. Additionally, the technology development will provide complementary cation and anion analysis of glycoconjugates to enable new dimensions for capturing complex modification states. Together these technologies will advance knowledge of how to leverage tandem mass spectrometry for glycoconjugate characterization, will generate new insights into the combinatorial epitopes that comprise the glycocode and will provide valuable opportunities for training academic and industrial researchers. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.