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

跨结构生物学和生物制药的原生自上而下质谱数据分析的综合解决方案

• 批准号: 10621751
• 负责人: Kenneth Durbin
• 金额: $ 79.67万
• 依托单位: PROTEINACEOUS, INC.
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621751
• 关键词: 3-Dimensional; Address; Adopted; Algorithms; Amino Acid Sequence; Antibodies; Binding; Bioinformatics; Biological Process; Clinical; Clip; Complex; Computer software; Cryoelectron Microscopy; Data; Data Analyses; Databases; Dedications; Development; Disease; Dissociation; Electron Microscopy; Elements; Ensure; Family; Grant; Heterogeneity; Individual; Industry; Informatics; Intuition; Ions; Learning; Ligand Binding; Ligands; Maps; Mass Spectrum Analysis; Modification; Molecular Weight; Names; Pathway Analysis; Peptides; Pharmaceutical Preparations; Phase; Protein Complex Subunit; Protein Databases; Proteins; Research; Scientist; Side; Software Tools; Structural Biologist; Techniques; Technology; Therapeutic Studies; Time; Validation; Visualization; Work; X-Ray Crystallography; algorithm development; commercialization; complex data; drug development; experience; experimental study; high throughput analysis; improved; industry partner; insight; mass spectrometer; novel; pre-clinical; protein complex; protein structure; screening; search engine; small molecule; stoichiometry; structural biology; success; therapeutic development; therapeutic protein; tool

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.

项目概要 原生自上而下质谱法正在继续发展成为一种强大的分析技术，可以 生成有关蛋白质形式和蛋白质复合物天然状态的转化数据。技术进步 近年来，质谱仪的应用推动了 nTDMS 的快速崛起，为 结构生物学研究和蛋白质疗法开发。例如，对非共价键的新见解 配体结合和定位可以通过与传统结构互补的 nTDMS 数据来揭示 生物学方法，例如电子显微镜和核磁共振。药物开发商可以应用 nTDMS 进行研究 临床前和临床环境中的治疗性蛋白质，以了解有关存在的修饰和修饰的更多信息 正在形成复合物。新型蛋白质和蛋白质复合物药物分子的数量不断增加， 每个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，取得了长足进步 已在大分子质量测定的算法开发方面取得进展；然而，没有完全 目前存在 nTDMS 特色分析平台，还集成了搜索。这里，蛋白质是 开发并商业化名为 ProSight Native 的新软件来填补这一分析空白。在第一阶段 该开发引入了第一个平台，用于分析来自 经典的 nTDMS 复合下降实验。该平台为完整的复合物及其它们提供解卷积 解离的亚基，以及搜索亚基鉴定和复杂的化学计量计算 成分测定。对于第二阶段的资助，将进行一系列新的研究，以实现 即将商业化的综合平台，具有改进的原生反卷积算法 proteoforms、首个 nTDMS 特异性高通量搜索以及强大的定量工作流程 生物制药应用。主要开发工作将用于 nTDMS 搜索的新技术 蛋白质形式和复合物，包括用于自动化学计量评分的多管齐下的方法 作业。此外，结构生物学元素将与蛋白质碎片数据一起整合 将这些重要的组成部分结合在一起并促进两者之间的联系。最后，结果将 显示在直观的、以结构生物学为中心的查看器中，使分析易于大众理解 光谱学家和非质谱学家都一样。总的来说，ProSight Native 的目标是显着推进 nTDMS 具有这些新颖的功能，同时还提高了对可生成的强大数据的可访问性 使用 nTDMS 技术。

项目成果

Advancing Intact Protein Quantitation with Updated Deconvolution Routines.

通过更新的解卷积程序推进完整蛋白质定量。

• 发表时间: 2023-10-10
• 影响因子: 7.4
• 作者: Robey, Matthew T;Utley, Daisha;Greer, Joseph B;Fellers, Ryan T;Kelleher, Neil L;Durbin, Kenneth R
• 通讯作者: Durbin, Kenneth R