Development of Next Generation Mass Spectrometric Instrumentation for Proteomics

开发下一代蛋白质组学质谱仪器

• 批准号: 10240528
• 负责人: Brian T Chait
• 金额: $ 70.91万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10240528
• 关键词: Address; Area; Biological; Biological Assay; Biology; Brain; Cancer Biology; Cells; Chemistry; Collection; Communicable Diseases; Complex; DNA; DNA sequencing; Detection; Development; Dideoxy Chain Termination DNA Sequencing; Dimensions; Disease; Fluorescence; Genome; Genomics; Goals; Hand; Human; Human Genome; Immunology; Infection; Ions; Location; Mass Spectrum Analysis; Measures; Methods; Mind; Modernization; Noise; Organism; Post-Translational Protein Processing; Process; Protein Analysis; Proteins; Proteome; Proteomics; Research; Research Personnel; Sampling; Signal Transduction; Specific qualifier value; Speed; Synapses; Techniques; Technology; Time; applied biomedical research; attenuation; cancer cell; cell type; comparative; cost; design; falls; genome sequencing; high throughput analysis; improved; innovation; instrumentation; interest; mass spectrometer; new technology; next generation; parallelization; pathogen; personalized medicine; protein crosslink; simulation; single cell analysis; structural biology; success; technological innovation; therapeutic development; tool; transcriptome; transcriptome sequencing; transcriptomics; wasting

Project Summary/Abstract Currently, most mass spectrometric (MS) analyses for proteomics are performed in sequential mode, wherein various species in a sample are selected and interrogated one after another. As a consequence of the finite time needed to examine each species in turn, sequential mode MS suffers from inescapable limitations in sensitivity, speed and ability to analyze all ions, especially when the composition of the ion beam is complex and rapidly changing. These limitations have kept vast tracts of biology and biomedicine, including, for example, deep single cell proteome analysis, out of reach of the current MS technology. In the present proposal, we posit that sensitivity, speed and dynamic range can be vastly improved by performing MS in parallel (by analogy to Next Generation DNA Sequencing), thus overcoming the technical barriers inherent to current commercial mass spectrometers that operate largely in sequential mode. Here, we propose to develop new MS instrumentation to execute MS in a massively parallel manner, with two major objectives in mind: Objective 1. Increase the sensitivity, speed and depth of proteome analyses by up to and ultimately beyond 1000-fold. The current sequential MS approaches can be likened to sampling the Niagara Falls with a bucket, where the majority of the sample is wasted. Our proposed parallel MS technology is designed to eliminate this immense waste. Objective 2. Filter noise in real time to eliminate unwanted ion background prior to MS analysis, thereby maximizing the MS utilization of the sample ions of interest and the resulting signal-to- noise ratios, as well as providing increased dynamic range to measure very low abundance components in the presence of highly abundant components. Successful attainment of these objectives will allow deep, comprehensive, high throughput analyses of proteomes in cases where sample availability is limiting, where the components of interest elude detection due to their low abundance, or when single cell analysis is needed to address the biological or biomedical question at hand. Success in this endeavor will propel forward many areas of basic and applied biomedical research that require proteomic analyses in a manner analogous to the immense progress that has been made through development of Next Generation DNA Sequencing.

项目概要/摘要 目前，大多数蛋白质组学质谱 (MS) 分析都是按顺序进行的 模式，其中样品中的各种物种被依次选择和询问。作为 依次检查每个物种所需的有限时间的结果，连续模式 MS 在分析所有离子的灵敏度、速度和能力方面不可避免地受到限制，尤其是 当离子束成分复杂且快速变化时。这些限制有 保存了大量的生物学和生物医学知识，包括例如深层单细胞蛋白质组 分析，目前的 MS 技术无法达到。在本提案中，我们认为 通过并行执行 MS（通过 类比下一代 DNA 测序），从而克服了固有的技术障碍 目前的商用质谱仪主要以顺序模式运行。 在这里，我们建议开发新的 MS 仪器来大规模并行执行 MS 方式，牢记两个主要目标： 目标 1. 将蛋白质组分析的灵敏度、速度和深度提高多达和 最终超过1000倍。当前的连续 MS 方法可以比作采样 用桶清洗尼亚加拉大瀑布，大部分样本都被浪费了。我们提出的并行 质谱技术旨在消除这种巨大的浪费。 目标 2. 在 MS 分析之前实时过滤噪声以消除不需要的离子背景， 从而最大限度地提高感兴趣的样品离子的 MS 利用率以及由此产生的信号到 噪声比，以及提供更大的动态范围来测量非常低的丰度 存在高度丰富的成分的成分。 成功实现这些目标将实现深入、全面、高通量 在样本可用性有限的情况下进行蛋白质组分析，其中 由于丰度低，或者需要进行单细胞分析时，兴趣会逃避检测 解决当前的生物学或生物医学问题。这项努力的成功将推动 推进许多需要蛋白质组学分析的基础和应用生物医学研究领域 类似于通过开发 Next 所取得的巨大进步 一代DNA测序。