The GCE4All Center: Unleashing the Potential of Genetic Code Expansion for Biomedical Research

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EQUIPMENT SUPPLEMENT ABSTRACT The GCE4All Biomedical Technology Development and Dissemination Center at Oregon State University (OSU) serves to optimize, develop, and broadly disseminate Genetic Code Expansion (GCE) technology – the engineering of cellular translation to express proteins containing non-canonical amino acids (ncAAs). During its envisioned lifespan of ≤15 years, the Center's mission is to effectively extend existing GCE technologies for facile use by non-specialists, and to broadly disseminate them via widespread education, effective training, and by providing sustainable access to optimized technologies via established repositories – enabling powerful GCE approaches to become standard, widely-used tools of biomedical researchers. As we build new GCE encoding systems, quantifying the intact mass of the ncAA-proteins is critical to verify the site-specific encoding of ncAAs, to verify that there is no loss in fidelity by mis-encoding of natural amino acids, and to evaluate biorthogonal labeling reactions on proteins. However, the center does not have equipment to characterize intact protein mass. The university MS facility has two MS systems capable of intact protein mass characterization, an old Waters Q- Tof MS system soon to be decommissioned and a high res-Q-TOF Lumos system. This facility unfortunately is not able to provide the daily or weekly access for analysis of intact protein mass needed by the center and they cannot accommodate walk up service from center staff. Here, we request funds to acquire a benchtop MS system, 6230B TOF MS system with ESI source and 1290 LCMS system with autosampler from Agilent. A center housed MS system capable of intact protein mass analysis would allow us to quickly evaluate proteins from the center and DBP labs. We expect the instrument will serve 50-100 researchers per year, including all personnel in the center, DBP collaborators and 2 workshops of 20 people per year. A permanent center staff member will be assigned to MS equipment maintenance and user training. The in-house ability to run and process our own intact protein MS analysis on a daily basis will greatly accelerate the center mission regarding GCE technology development, optimization and dissemination for DBPs and GCE trainees. The summary of our original grant is included in our research strategy document.