Enable ultra-high throughput, ultra-sensitive and more quantitative broad proteomics measurements from much smaller samples

能够从更小的样品中进行超高通量、超灵敏和更定量的广泛蛋白质组学测量

• 批准号: 10461819
• 负责人: RICHARD D SMITH
• 金额: $ 50.63万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10461819
• 关键词: Address; Automobile Driving; Bioinformatics; Biological; Biomedical Research; Communities; Complement; Coupled; Data; Data Analyses; Development; Evaluation; Fluorescence-Activated Cell Sorting; Goals; Heart; Informatics; Ions; Isomerism; Laboratories; Mass Spectrum Analysis; Measurement; Methodology; Nature; Peptides; Performance; Property; Proteins; Proteome; Proteomics; Reproducibility; Research; Resolution; Resource Development; Resources; Retinal blind spot; Role; Sample Size; Sampling; Shapes; Speed; Stable Isotope Labeling; Structure; Techniques; Technology; Time; Work; base; bioinformatics tool; commercialization; community engagement; computerized data processing; computerized tools; data quality; improved; insight; ion mobility; laser capture microdissection; liquid chromatography mass spectrometry; novel; tool development; ultra high resolution

Project Summary – TR&D 2 The Resource has the overall goal of broadly impacting biomedical research by providing the abilities to: obtain high quality proteomics data from much smaller samples, produce more quantitative and comprehensive measurements, generate improved and more extensive information on low abundance components, distinguish presently problematic peptide isomers, and enable the study of much larger sample sets than presently practical by providing increases in measurement sensitivity and throughput. The technology efforts of TR&D 2 will enable these capabilities by providing high resolution separations and ion manipulations with ion mobility- mass spectrometry that result in proteomics measurements with much greater sensitivity and throughput, which in conjunction with the `front-end' developments under TR&D 1, will provide the basis for broadly effective applications of proteomics from much smaller samples. TR&D 2 will develop new platforms based on our novel Structures for Lossless Ion Manipulations (SLIM) technology. These platforms will provide ultra-fast and ultra-high resolution ion mobility (IM) separations that will complement the distinctive advantages provided by both fast time-of flight MS and ultra-high mass accuracy and resolution FTMS (specifically Orbitrap-based MS) platforms. This work will benefit from the inherently fast, robust, and highly reproducible nature of ultra-high resolution IM separations, as well as the distinctive peptide/protein size/shape information (collision cross sections) provided by IM. In combination with the bioinformatics developments of TR&D 3, TR&D 2 will enable improved quantification and proteome coverage in conjunction with high throughput measurements. Overall, TR&D 2 will enable: (1) much more extensive proteomic information from extremely small samples (e.g. from Fluorescence-activated cell sorting and laser capture microdissection) when applied in conjunction with the developments of TR&D 1; (2) large gains in proteomic throughput (measurement speeds) based upon fast ultrahigh-resolution IM separations and related MS-based advances; (3) more quantitative and higher data quality measurements that effectively provide a convergence of the attractive properties of global and targeted MS-based proteomic measurements; and (4) broader proteome measurement coverage that also addresses important `blind spots' of current technologies, such as presently indistinguishable PTMs and other peptide isomers. These efforts will build upon previous Resource developments and will be facilitated by key computational and bioinformatics tool developments under TR&D 3. In combination, these efforts will provide a basis for rapid implementation and initial evaluation of the new capabilities in a set of challenging biomedical projects, as well as their effective dissemination to the research community, including seeding the technology in several outside laboratories and facilitating their initial commercialization, so as to assure broad and sustained availability.

项目摘要-研发2 该资源的总体目标是通过提供以下能力来广泛影响生物医学研究： 从小得多的样本中获得高质量的蛋白质组学数据，产生更多定量和全面的数据 测量，生成关于低丰度组分的改进和更广泛的信息，区分 目前有问题的多肽异构体，并使研究比目前大得多的样本集成为可能 通过提高测量灵敏度和吞吐量，实现了实用化。R&D 2的技术努力 将通过提供具有离子迁移率的高分辨率分离和离子操纵来实现这些能力- 导致蛋白质组学测量具有更高的灵敏度和吞吐量的质谱学， 与研发1项下的“前端”发展相结合，将提供广泛的基础 从小得多的样本中有效地应用蛋白质组学。研发2将基于以下方面开发新平台 我们的无损离子操纵(SLIM)技术的新结构。这些平台将提供超快的 和超高分辨率离子迁移率(IM)分离，这将补充独特的优势 由快速飞行时间MS和超高质量精度和分辨率FTMS(特别是 基于Orbitrap的MS)平台。这项工作将受益于固有的快速、健壮和高度可重复性 超高分辨率IM分离的性质，以及独特的肽/蛋白质大小/形状 IM提供的信息(碰撞横截面)。结合生物信息学的发展 研发3、研发2将实现更好的量化和蛋白质组覆盖，与高 吞吐量测量。总体而言，研发2将使：(1)更广泛的蛋白质组信息来自 极小的样本(例如，来自荧光激活细胞分选和激光捕获显微切割) 与研发结合应用1；(2)蛋白质组学产量的大幅提高 (测量速度)基于快速超高分辨率IM分离和相关的基于MS的进展； (3)更多量化和更高数据质量的衡量标准，有效地提供 基于整体和靶向MS的蛋白质组测量的吸引人的特性；以及(4)更广泛的蛋白质组 测量覆盖面，也解决了当前技术的重要“盲点”，例如目前 难以区分的PTMS和其他多肽异构体。这些工作将建立在以前资源的基础上 开发，并将通过以下项目下的关键计算和生物信息学工具开发来促进 研发3.结合起来，这些努力将为快速实施和初步评估 一系列具有挑战性的生物医学项目中的新能力，以及它们有效地传播到 研究界，包括在几个外部实验室播种技术并促进他们的 初步商业化，以确保广泛和可持续的供应。