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

项目概要/摘要（30行） 该项目旨在解决目前快速和全面（大规模， 结构）剖析生物大分子。而高分辨率离子迁移率和质谱 虽然分离技术已经得到了广泛的发展，但它们的应用在很大程度上局限于 小分子（m/z < 2000）。以高吞吐量访问结构和大量信息的能力 大生物分子的时尚是质谱分析中的一个理想目标。为了实现这一目标， 我们将开发一种基于无损结构的高分辨率离子迁移率化学分析装置 离子操作，并与质谱联用，获得高通量高分辨率 生物大分子的IMS/MS。生物大分子中显著异质性的研究 系统，即，生物大分子，生物研究的关键将通过发展 一种新的高分辨率、高通量分析系统。SLIM平台目前已启用， 超高分辨率离子迁移率分布（直到分离同位素异构体和同位素物的程度） 并且小分子离子（<3000道尔顿）的操作将被优化以有效地限制， 运输和分离生物大分子。SLIM的初始优化将通过耦合 用飞行时间质谱系统SLIM-TOF将用于执行高分辨率IMS/MS的集合， 大生物分子种类（具有由TOF规定的约20，000 Da的质量上限）。我们将 使用基于SLIM的IMS分离和其他操作来揭示广泛的结构 生物大分子的异质性。此外，我们还将进行规模和质量选择性软着陆， 大生物分子的动态切换和重新路由（沉积基板）的选择性 SLIM中的离子迁移率分布。软着陆物种将使用cryo-EM和 互补的成像技术，以阐明大生物分子的形态， 迁移率或碰撞截面。离子迁移率信息和成像的结合将 提供了大生物分子的详细结构表征。

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.