TR&D 5: Top-Down for the Masses

技术研发项目5：自上而下服务大众 摘要 对于使用完整蛋白质作为质谱法中的主要测量单位存在强有力的论据- 基于蛋白质组学蛋白质型假说，这表明它是特定的蛋白质型， 与表型相关，与从事转化和基础科学的研究人员产生了强烈的共鸣。在 在上一个资助期，NRTDP为发现科学开发了有效的定量工作流程，并应用于 这些工作流程到无数的生物系统。为了认识到我们有责任促进更广泛的社区，我们 选择创建一个完整的TR&D，专注于使我们的定量工作流程足够强大， 通过社区参与导出到非专家实验室。要做到这一点，每一步， 工作流程，从样品制备和分离到质谱检测和数据分析， 创新以提高鲁棒性。新的样品制备技术，加上更强大，更高的流量 色谱法将减轻与纳米流色谱法相关的问题。更清洁的样品加上现场 不对称离子迁移谱（FAIMS）也将改善质谱仪的正常运行时间。除了 提高硬件的鲁棒性，资源也将发布更新的软件工具，包括AUTOPILOT ACQUIRE、TDPORTAL和TDVIEWER，包括简化的定量数据采集、处理和 分析工作流。这些创新的结合将使资源能够有效地出口 这些实验是在内部开发的，社区对执行实验的兴趣越来越大， 都是蛋白质分解的

Technology Research and Development Project 5: Top-Down for the Masses Abstract Strong arguments exist for utilizing intact proteins as the primary unit of measurement in mass spectrometry- based proteomics. The proteoform hypothesis, which suggests that it is specific proteoforms that are most tightly linked to phenotype, resonates strongly with researchers performing both translational and basic science. In the previous granting period, the NRTDP developed potent quantitative workflows for discovery science and applied these workflows to myriad biological systems. In recognition of our duty to foster a broader community, we have chosen to create an entire TR&D focused on making our quantitative workflows robust enough that they can be exported to non-expert labs through Community Engagement. To accomplish this, each step in these workflows, from sample preparation and separation to mass spectrometric detection and data analysis will be innovated to improve robustness. New sample preparation technology coupled with more robust, higher flow chromatography will alleviate issues related to nanoflow chromatography. Cleaner samples coupled with field asymmetric ion mobility spectrometry (FAIMS) will also improve mass spectrometer up-time. In addition to improved hardware robustness, the resource will also publish updated software tools, including AUTOPILOT ACQUIRE, TDPORTAL and TDVIEWER, which include streamlined quantitative data acquisition, processing, and analysis workflows. The combination of these innovations will give the resource the capability to efficiently export the experiments developed in-house to a community that has a growing appetite for performing experiments that are proteoform-resolved.