Multiplexed Charge Detection Mass Spectrometer for Extended Mass and Collisional Cross Section Measurements

用于扩展质量和碰撞截面测量的多重电荷检测质谱仪

基本信息

批准号：
10267735
负责人：
Evan R Williams
金额：
$ 30.18万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-22 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10267735
关键词：
Area Biological Bypass Cell physiology Charge Complex Complex Mixtures Consumption Data Detection Development Dissociation Drug Targeting Electrospray Ionization Electrostatics Event Frequencies Gases Generations Goals Health Heterogeneity Hour Human Individual Injections Ions Lipoproteins Macromolecular Complexes Mass Spectrum Analysis Measurement Measures Methods Modeling Molecular Optics Parents Pathway interactions Phase Polymers Process Proteins Publishing Research Resolution Sampling Scheme Signal Transduction Solvents Speed Structure System Techniques Testing Therapeutic Therapeutic Agents Time Viral Vector Virus-like particle Work base design detector evaporation experience experimental study improved innovation insight instrument instrumentation ion mobility macromolecular assembly macromolecule mass spectrometer molecular assembly/self assembly molecular shape next generation novel prevent protein aggregation stem targeted delivery tool transmission process viral DNA

项目摘要

Project Summary Large biomolecular assemblies constitute much of the molecular machinery necessary for cellular function. These assemblies are often the target of a host of therapeutic molecules. Other large biomolecular assemblies, including virus-like particles, proteinaceous cellular compartments, and some large synthetic polymers, are potential delivery agents for these therapeutics. The large molecules and molecular assemblies involved in cellular function can be challenging to analyze using conventional mass spectrometry methods owing to their high mass (>MDa) and heterogeneity. Electrospray ionization can transfer intact large molecules/complexes into the gas phase, but overlapping and unresolved charge states that stem from the heterogeneity inherent to high mass analytes often prevent mass measurements using conventional mass spectrometers. Charge detection mass spectrometry weighs individual ions, avoiding these interferences between ions and can be used to analyze molecules with masses well above 100 MDa. However, this method can be slow because only one ion is analyzed at a time. Here, a new generation of charge detection mass spectrometry is proposed which provides information about the mass, the collisional cross section and the dissociation pathways of individual ions of large macromolecular complexes, information that cannot be obtained using conventional instruments. A key innovation is the development of multiplexing methods that make it possible to simultaneously weigh many individual ions. These methods have the potential to increase the speed of these measurements by up to 150x and reduce sample analysis times to less than one minute. This ion multiplexing is aided by a novel decoupling scheme whereby ions with a controlled range of energies are introduced into the electrostatic ion detector trap so that two or more ions with the same m/z can have different frequencies. The mass of each ion can be obtained from simultaneous measurements of the individual ion frequency, energy, and charge. This scheme significantly reduces the potential for overlapping ion signal by distributing the signal over a broad frequency bandwidth. The rate of ion energy change can also be determined from these measurements, providing information about collisional cross sections and giving insight into molecular shape. Similarly, individual ion fragmentation events can be tracked and used to obtain additional structural information.

项目概要大型生物分子组装体构成了许多必需的分子机器细胞功能。这些组装体通常是许多治疗分子的目标。其他大型生物分子组装体，包括病毒样颗粒、蛋白质细胞隔室和一些大型合成聚合物是这些物质的潜在输送剂疗法。参与细胞功能的大分子和分子组装体可以是由于其质量较高，使用传统的质谱方法进行分析具有挑战性 (>MDa) 和异质性。电喷雾电离可以完整地转移大分子/复合物进入气相，但重叠和未解析的电荷表明源于高质量分析物固有的异质性通常会妨碍质量测量使用传统的质谱仪。电荷检测质谱法称重个体离子，避免了离子之间的这些干扰，可用于分析分子质量远高于 100 MDa。然而，这种方法可能很慢，因为只有一个离子一时分析。这里提出了新一代电荷检测质谱法它提供有关质量、碰撞截面和解离的信息大分子复合物的单个离子的路径，这些信息是无法被使用常规仪器获得。一项关键的创新是多路复用技术的发展可以同时称量许多单独离子的方法。这些方法有潜力将这些测量的速度提高多达 150 倍并减少样品分析时间缩短至不到一分钟。这种离子多路复用是由一种新颖的去耦方案，将具有受控能量范围的离子引入到静电离子检测器陷阱，使两个或多个具有相同 m/z 的离子可以具有不同的频率。每个离子的质量可以通过同时测量单个离子的频率、能量和电荷。该方案显着降低了通过将信号分布在较宽的频率带宽上来重叠离子信号。的比率离子能量变化也可以从这些测量中确定，提供信息关于碰撞横截面并深入了解分子形状。同样，单个离子可以跟踪碎片事件并使用它来获取附加的结构信息。