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

设备配置摘要 俄勒冈州州立大学（OSU）的GCE 4 All生物医学技术开发和传播中心 用于优化，开发和广泛传播遗传密码扩展（GCE）技术- 工程化细胞翻译以表达含有非典型氨基酸（ncAA）的蛋白质。期间 该中心的使命是有效地扩展现有的GCE技术， 非专业人员使用，并通过广泛的教育、有效的培训和 通过已建立的知识库提供对优化技术的可持续访问-实现强大的GCE 方法成为生物医学研究人员的标准，广泛使用的工具。当我们构建新的GCE编码时， 系统，定量ncAA蛋白的完整质量对于验证ncAA的位点特异性编码至关重要， 为了验证天然氨基酸的错误编码没有损失保真度，并评估双正交 蛋白质上的标记反应。然而，该中心没有设备来表征完整的蛋白质质量。 该大学的MS设施有两个MS系统能够完整的蛋白质质量表征，一个旧的沃茨Q- TOF MS系统即将退役，而高分辨率-Q-TOF Lumos系统。不幸的是， 不能提供中心所需的完整蛋白质质量的每日或每周分析， 不能接受中心工作人员的步行服务。在这里，我们要求资金购买台式MS系统， 具有ESI源的6230 B TOF MS系统和来自Agilent的具有自动进样器的1290 LCMS系统。一个中心， 能够进行完整蛋白质质量分析的MS系统将使我们能够快速评估来自中心的蛋白质 DBP实验室我们预计该仪器每年将为50-100名研究人员提供服务，包括研究中心的所有人员。 中心，DBP合作者和每年20人的2个研讨会。一个永久的中心工作人员将是 分配到MS设备维护和用户培训。内部运行和处理我们自己的完整能力 每天进行蛋白质质谱分析，将大大加快该中心在GCE技术方面的使命 为DBP和GCE受训人员制定、优化和传播。 我们的原始补助金摘要包含在我们的研究战略文件中。

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.