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

项目摘要/摘要(30行) 该项目旨在解决当前在快速和全面(海量、 结构)生物大分子的剖析。而高分辨率离子迁移率和质谱学 分离技术得到了广泛的发展，其应用在很大程度上局限于 小分子(m/z&lt；2000)。能够以高吞吐量访问结构和海量信息 大生物分子的流行是质谱学的一个令人向往的目标。为了实现这一目标， 我们将开发一种基于结构无损的高分辨率离子迁移率化学分析装置 离子操作，并与质谱学相结合，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 对生物学研究至关重要的系统，即大生物分子，将通过 一种新的高分辨率、高通量分析系统。超薄平台，目前支持 超高分辨率离子迁移率剖面图(可用于分离同位素异构体和同位素) 对小分子离子(3000道尔顿)的操作将得到优化，以有效地限制， 运输和分离大分子生物分子。SLIM的初始优化将通过耦合进行 使用TOF-MS系统。SLIM-TOF将用于执行高分辨率的IMS/MS系综 大生物分子物种(TOF规定的质量上限约为20000 Da)。我们会 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，我们将进行大小和质量选择性软着陆 利用选择性的动态切换和重路由(到沉积基质)的大生物分子 离子迁移率在薄层中的分布。软着陆物种将使用低温EM和 补充成像技术以阐明选定的大生物分子的形态 迁移率或碰撞横截面。离子迁移率信息和成像的结合将 提供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.