A comprehensive solution for native top down mass spectrometry data analyses across structural biology and biopharma

项目总结 自然自上而下的质谱学正在继续发展成为一种强大的分析技术，它可以 生成关于蛋白质形式和蛋白质复合体天然状态的变革性数据。技术进步 近年来，质谱仪的出现刺激了nTDMS的迅速崛起，为 结构生物学研究和蛋白质疗法的发展。例如，对非共价的新见解 NTDMS数据可以揭示配体的结合和定位，这些数据是对传统结构的补充 电子显微镜和核磁共振等生物学方法。药物开发者可以应用nTDMS进行研究 临床前和临床环境中的治疗性蛋白质，以了解更多现有的修饰和 正在形成的复合体。新型蛋白质和蛋白质复合药物分子的数量不断增加， 每一项都面临着一系列困难的分析和数据分析挑战。在nTDMS的生物信息学方面，Strides 已经在大分子质量测定的算法开发中取得了进展；然而，还没有完全 目前存在的nTDMS的特色分析平台也集成了搜索。在这里，蛋白质是 开发并商业化一款名为ProSight Native的新软件，以填补这一分析空白。在第I阶段， 该开发是首个此类平台，用于分析来自 经典的nTDMS络合降压实验。该平台为完整的复合体和它们的 解离的亚基，以及搜索亚基识别和络合物的化学计量计算 成分测定。对于第二阶段的拨款，将进行一系列新的研究，以使 将商业化的综合平台，采用改进的本机反卷积算法 ProteoForms，有史以来第一个特定于nTDMS的高通量搜索，以及强大的量化工作流 生物制药应用。主要开发工作将在此花费在nTDMS搜索的新技术上 蛋白质形式和复合体，包括多管齐下的自动化学计量计分方法 任务。此外，结构生物学元素将与蛋白质形式碎片数据一起整合 将这些重要组成部分结合在一起，促进两者之间的联系。最后，结果将是 在直观的、以结构生物学为中心的查看器中显示，这将使大量分析变得容易 光谱学家和非质谱学家一样。总体而言，ProSight Native的目标是大幅提升 NTDM具有这些新颖的功能，同时还提高了对可以生成的强大数据的可访问性 使用nTDMS技术。

PROJECT SUMMARY Native top down mass spectrometry is continuing to evolve into a powerhouse analytical technique that can generate transformative data on the native state of proteoforms and protein complexes. Technological advances in mass spectrometers have spurred the rapid rise of nTDMS over recent years, paving impressive avenues for structural biology research and protein therapeutics development. For instance, new insights into non-covalent ligand binding and localization can be revealed by nTDMS data that are complementary to traditional structural biology approaches such as electron microscopy and NMR. Drug developers can apply nTDMS to study therapeutic proteins in preclinical and clinical settings to learn more about modifications that are present and the complexes being formed. The number of novel protein and protein complex drug molecules is ever increasing, each with a host of difficult analytical and data analysis challenges. On the bioinformatics side of nTDMS, strides have been made in algorithmic development for mass determination of large molecules; however, no fully featured analysis platform for nTDMS currently exists that also integrates search. Here, Proteinaceous is developing and commercializing a new software named ProSight Native to fill this analysis void. In Phase I of the development, a first-of-its-kind platform was introduced for analyzing targeted protein complex data from the classic nTDMS complex-down experiment. The platform offers deconvolution for intact complexes and their dissociated subunits, as well as search for subunit identification and a stoichiometry calculation for complex composition determination. For the Phase II grant, a host of new research will be undertaken to enable a comprehensive platform to be commercialized that features an improved deconvolution algorithm for native proteoforms, the first-ever nTDMS-specific high-throughput search, and robust quantitation workflows for biopharma applications. Major development effort will be spent here on new techniques for nTDMS searches of both proteoforms and complexes, including a multi-pronged approach for scoring automated stoichiometry assignments. Additionally, structural biology elements will be integrated alongside proteoform fragmentation data to bring these important components together and facilitate connections between the two. Lastly, the results will be displayed in an intuitive, structural biology-centric viewer that will make analysis approachable for mass spectrometrists and non-mass spectrometrist alike. In total, ProSight Native will aim to significantly advance nTDMS with these novel features, while also increasing accessibility to the powerful data that can be generated using nTDMS techniques.