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.

项目摘要/摘要 当前，大多数蛋白质组学的质谱分析（MS）分析是在顺序的 模式，其中选择样品中的各种物种并接一个地审问。作为 依次检查每个物种所需的有限时间的结果MS 敏感性，速度和分析所有离子的能力不可避免的限制，尤其是 当离子束的组成复杂且快速变化时。这些限制有 保持大量的生物学和生物医学，包括例如深单细胞蛋白质组 分析，无法实现当前的MS技术。在本提案中，我们认为 通过并行执行MS（通过 类似于下一代DNA测序），因此克服了 当前的商业质谱仪在很大程度上以顺序模式运行。 在这里，我们建议开发新的MS仪器以在大规模平行的情况下执行MS 方式，有两个主要目标： 目标1。提高蛋白质组分析的敏感性，速度和深度。 最终超过1000倍。当前的顺序MS方法可以比喻为采样 尼亚加拉（Niagara）倒下了一个水桶，大部分样品都浪费了。我们提出的平行 MS Technology旨在消除这种巨大的废物。 目标2。实时的过滤噪声，以消除MS分析之前消除不需要的离子背景， 从而最大程度地利用了感兴趣的样品离子的MS利用率，并产生的信号对 - 噪声比，并提供增加动态范围以测量非常低的丰度 组件存在高度丰富的组件。 成功实现这些目标将允许深刻，全面，高吞吐量 在样本可用性限制的情况下，对蛋白质组的分析，其中的成分 兴趣避免检测由于其较低的丰度或需要单细胞分析而引起的检测 解决眼前的生物学或生物医学问题。这项努力的成功将推动 转发基本和应用生物医学研究的许多领域，需要在 类似于通过发展下一步取得的巨大进步的方式 生成DNA测序。