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

项目概要- TR&D 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： 更小样本的高质量蛋白质组学数据， 测量，产生关于低丰度组分的改进的和更广泛的信息， 目前有问题的肽异构体，并使研究更大的样品集比目前 通过提供测量灵敏度和吞吐量的增加而实用。TR&D 2的技术成果 将通过提供高分辨率分离和具有离子迁移率的离子操作来实现这些能力- 质谱法导致具有更高灵敏度和通量的蛋白质组学测量， 与TR&D 1下的“前端”开发相结合，将为广泛的 从更小的样本中有效地应用蛋白质组学。TR&D 2将开发基于 我们的新型无损离子操纵结构（SLIM）技术。这些平台将提供超快的 和超高分辨率离子迁移率（IM）分离，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（具体地， 基于Orbitrap的MS）平台。这项工作将受益于固有的快速，稳健和高度可重复性 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， TR&D 3，TR&D 2将能够提高定量和蛋白质组覆盖率， 吞吐量测量。总的来说，TR&D 2将使：（1）更广泛的蛋白质组学信息， 极小的样品（例如，来自荧光激活细胞分选和激光捕获显微切割）， 与TR&D 1的开发结合使用;（2）蛋白质组通量的大幅增加 （测量速度）的基础上快速超高分辨率IM分离和相关的MS为基础的进步; (3)更定量和更高的数据质量测量，有效地提供了收敛的 基于MS的全局和靶向蛋白质组学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 不可区分的PTM和其他肽异构体。这些努力将建立在以前的资源基础上。 开发，并将通过以下关键计算和生物信息学工具的开发来促进 3.第三章这些努力结合在一起，将为迅速执行和初步评价 在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 研究社区，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，以确保广泛和持续的可用性。

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.