HIGHLY SELECTIVE MACROBIOMOLECULAR ISOLATION, SOFT-LANDING AND CHARACTERIZATION USING STRUCTURES FOR LOSSLESS ION MANIPULATIONS

使用无损离子操作结构进行高选择性大生物分子分离、软着陆和表征

• 批准号: 10713577
• 负责人: Sandilya Garimella
• 金额: $ 40.66万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713577
• 关键词: Address; Biological; Biological Phenomena; Biology; Chemicals; Collection; Complex; Coupled; Coupling; Cryoelectron Microscopy; Dendrimers; Deposition; Development; Devices; Gases; Goals; Heterogeneity; Image; Imaging Techniques; Ions; Mass Spectrum Analysis; Measurement; Methods; Molecular; Morphology; Property; Proteins; Resolution; Retrieval; Route; Shapes; Spectrometry; Structure; System; Techniques; Technology; Translating; Travel; aspirate; biological research; biological systems; biomacromolecule; dalton; electric field; ion mobility; macromolecule; nanoparticle; novel; particle; preservation; pressure; protein complex; simulation; small molecule; ultra high resolution

PROJECT SUMMARY/ABSTRACT (30 lines) This project aims to address the current gap in technology for rapid and comprehensive (mass, structure) profiling of biomacromolecules. While high resolution ion mobility and mass spectrometry separations techniques have been extensively developed, their application has been largely limited to small molecules (m/z < 2000). The ability to access structure and mass information in a high-throughput fashion for macrobiomolecules is an aspirational goal in mass spectrometry. In pursuance of this goal, we will develop a high-resolution ion mobility chemical analysis device based on Structures for Lossless Ion Manipulations, and in conjunction with mass spectrometry obtain high-throughput high-resolution IMS/MS of biomacromolecules. The study of the significant heterogeneity in large biomolecular systems, i.e., macrobiomolecules, critical to biological research will be enabled by the development of a new high resolution, high throughput analytical system. The SLIM platform, currently enabled for ultra-high resolution ion mobility profiling (up to the point of separating isotopomers and isotopologues) and manipulations of small molecular ions (<3000 Daltons) will be optimized to efficiently confine, transport and separate macrobiomolecules. Initial optimization of the SLIM will be done by coupling with a TOF-MS system. SLIM-TOF will be used to perform high resolution IMS/MS of ensembles of macrobiomolecular species (with an upper mass limit of about 20,000 Da dictated by the TOF). We will use SLIM-based IMS separations and other manipulations to reveal the extensive structural heterogeneity of macrobiomolecules. Further, we will perform size and mass selective soft-landing of macrobiomolecules using the dynamic switching and rerouting (to deposition substrates) of a selective ion mobility distribution in the SLIM. The soft-landed species will be characterized using cryo-EM and complimentary imaging techniques to elucidate the morphologies of macrobiomolecules with selected mobilities or collision-cross-sections. The combination of ion mobility information and imaging will provide a detailed structural characterization of macrobiomolecules.

项目摘要/摘要（30行） 该项目旨在解决当前技术差距，以实现快速和全面的差距（质量， 结构）生物分子的分析。而高分辨率离子迁移率和质谱法 分离技术已经广泛开发，它们的应用在很大程度上限于 小分子（m/z <2000）。在高通量中访问结构和质量信息的能力 大分子分子的时尚是质谱中的理想目标。为了实现这一目标， 我们将基于无损的结构开发高分辨率的离子迁移率化学分析设备 离子操作，并与质谱法结合获得高通量高分辨率 IMS/MS的生物大分子。大分子大分子的显着异质性的研究 系统，即大分子，对生物学研究至关重要 一个新的高分辨率高吞吐量分析系统。 Slim平台，当前启用了 超高分辨率离子迁移率分析（直到分离同位素和同位素分离） 小分子离子（<3000 daltons）的操作将被优化，以有效地局限 运输和分离大分子。 Slim的初始优化将通过耦合完成 使用TOF-MS系统。 Slim-TOF将用于执行高分辨率IMS/MS的合奏 大分子分子物种（大约20,000个DA由TOF决定）。我们将 使用基于细长的IMS分离和其他操纵来揭示广泛的结构 大分子的异质性。此外，我们将执行尺寸和质量选择性软地面 选择性的动态切换和重新布线（至沉积基板）的大分子 离子迁移率分布在细长中。软地面物种将使用冷冻EM和 与选定的大分子的形态阐明了免费成像技术 迁移率或碰撞区域。离子移动信息和成像的组合将 提供大分子的详细结构表征。