Developing High-Resolution Ion Mobility Spectrometry-Charge Detection-Mass Spectrometry for Rapid Analysis in the Megadalton to Gigadalton Regime

开发高分辨率离子淌度谱-电荷检测-质谱法，以实现兆道尔顿到千兆道尔顿范围内的快速分析

• 批准号: 10061629
• 负责人: DAVID E. CLEMMER
• 金额: $ 48.77万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-15 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10061629
• 关键词: Address; Amplifiers; Antibodies; Area; Atherosclerosis; Binding; Capsid; Cardiovascular Diseases; Cause of Death; Charge; Complement; Complex; Complex Mixtures; Computer Simulation; Coupled; Data; Data Collection; Detection; Development; Diagnostic tests; Ensure; Fractionation; Gene Transduction Agent; Genetic Load; Genetic Materials; Genome; Health; High Density Lipoproteins; Hour; Human; Individual; Ions; Lead; Lipoproteins; Low-Density Lipoproteins; Mass Spectrum Analysis; Measurement; Measures; Molecular; Molecular Weight; Noise; Pathogenesis; Pattern; Pharmaceutical Preparations; Plasma; Play; Research; Resolution; Role; Sampling; Spectrometry; Speed; Structural defect; Structure; System; Technology; Testing; Therapeutic; Time; Tube; Variant; Very low density lipoprotein; Viral Genes; Viral Proteins; Virion; Virus; Virus Assembly; Work; analytical method; base; cardiovascular disorder risk; cardiovascular risk factor; design; detector; gene therapy; improved; instrument; ion mobility; new therapeutic target; novel; novel strategies; programs; protein complex; prototype; tool; transmission process; whole genome

Project Summary This project will advance a new mass spectrometry (MS) technology, charge detection mass spectrometry (CDMS) for analyzing large biomolecular assemblies in the 10 to 100 nm size range (i.e., having molecular weights of ~1 MDa to 1 GDa). Species in this size range, such as viruses and lipoproteins, play critical roles in human health. However, they are difficult to detect and characterize. Conventional MS instruments can determine masses and fragmentation patterns of many types of biomolecules, including assemblies; but, such measurements are limited to species below ~1 MDa. In CDMS, the masses of individual ions are directly determined from simultaneous measurement of each ion’s mass-to-charge ratio and charge. Proof-of-principle studies using our CDMS prototype, show that accurate masses can be determined into the MDa to GDa regime. As shown in this proposal, this is an enabling advance. However, our prototype design has a limited mass resolving power (the maximum we have measured is m/Δm ~ 330). And, the time required for measuring a complete spectrum makes this instrument impractical for routine analyses. This project describes advances that will improve both the CDMS resolving power and spectrum acquisition speed – each by at least an order of magnitude. This will allow high-resolution mass spectra for large species to be routinely recorded for the first time. We will also develop an ion mobility spectrometry (IMS) interface for CDMS. The IMS separation will improve the overall peak capacity and simplify the analysis of heterogeneous mixtures by CDMS analysis. In addition, the mobility of an ion depends on its structure and thus provides information that complements the charge and mass information from CDMS. The combined high-resolution IMS-CDMS instrument will provide a powerful new approach for the analysis of large biomolecules. Once calibrated and tested, the IMS-CDMS instrument will be used to investigate numerous problems important to human health. These include: virus assembly and disassembly, the characterization gene therapy vectors, drug and antibody binding to viruses, and the identification of lipoprotein subclasses. Such measurements have transformative potential. Viral gene therapy vectors, for example, are difficult to characterize because of their large sizes and because the genetic material is contained within virus capsids. Critical issues, such as whether capsids contain the full genome, partial genomes, or no genome, must be addressed. High-resolution IMS-CDMS measurements will provide information about small structural defects, which are difficult (if not impossible) to detect by other means. Similarly, IMS-CDMS analyses will impact other areas, such as cardiovascular diseases - the leading cause of death in the US. Plasma lipoproteins play a key role in the main underlying cause, atherosclerosis. However, the major classes, HDL, LDL, and VLDL, all exist as broad distributions of sizes and compositions. Delineation of subclasses will enable development of more reliable diagnostic tests and better therapeutics. Preliminary results indicate that high-resolution IMS-CDMS will be able to resolve key lipoprotein subclasses.

项目摘要 该项目将推进新的质谱（MS）技术，电荷检测质谱法 （CDMS）用于分析10至100 nm尺寸范围内的大型生物分子组件（即具有分子 〜1 MDA至1 GDA的权重）。在这种尺寸范围内的物种，例如病毒和脂蛋白，在 人类健康。但是，它们很难检测和表征。传统的MS仪器可以 确定许多类型的生物分子的质量和碎片模式，包括组装；但是，这样 测量仅限于〜1 MDA以下的物种。在CDM中，单个离子的质量直接 由简单测量每个离子的质量和电荷的测量。原理证明 使用我们的CDMS原型的研究表明，可以确定准确的质量到GDA中 政权。如本提案所示，这是一个有利的进步。但是，我们的原型设计有限 质量分辨能力（我们测量的最大值为m/Δm〜330）。而且，测量所需的时间 完整的频谱使该仪器不切实际地进行常规分析。该项目描述了进步 这将提高CDM的分辨能力和频谱采集速度 - 至少要订单 数量级。这将允许在第一个常规记录大型物种的高分辨率质谱 时间。我们还将为CDMS开发一个离子迁移率（IMS）界面。 IMS分离将 通过CDMS分析来改善总体峰值能力并简化异质混合物的分析。在 此外，离子的移动性取决于其结构，因此提供了完成的信息 来自CDM的充电和质量信息。组合的高分辨率IMS-CDMS仪器将提供 强大的新方法来分析大型生物分子。校准和测试后，IMS-CDMS 仪器将用于调查对人类健康重要的许多问题。这些包括：病毒 组装和拆卸，表征基因治疗载体，药物和与病毒结合的抗体， 以及脂蛋白亚类的鉴定。这样的测量具有变革潜力。病毒基因 例如 材料包含在病毒衣壳中。关键问题，例如衣壳是否包含完整的基因组， 必须解决部分基因组或没有基因组。高分辨率IMS-CDMS测量将提供 有关小结构缺陷的信息，这些缺陷很难（如果不是不可能）通过其他方式检测。 同样，IMS -CDMS分析将影响其他领域，例如心血管疾病 - 主要原因 在美国死亡。血浆脂蛋白在主要根本原因（动脉粥样硬化）中起关键作用。然而， HDL，LDL和VLDL的主要类别都是大小和组成的广泛分布。描述 子类别将能够开发更可靠的诊断测试和更好的治疗。初步的 结果表明，高分辨率IMS-CDMS将能够解决关键的脂蛋白亚类。