High Resolution High Throughput Proteomics Platform for Cancer Research

项目总结/摘要 本项目的目标是评估一种新的高通量和超灵敏的平台 这将使更有效和更高通量的蛋白质组学测量， 推进癌症研究。新平台将提供高得多的分辨率离子 迁移谱（IMS）分离比目前的平台。这项工作还利用了 通过将这些新功能与更多 灵敏、高效的离子源和高精度的质谱技术。IMS的分辨率 实现将大大增加通过使用电力行波施加到一个非常长的 路径设计基于无损离子操纵结构（SLIM），同时还能够 要达到很高的灵敏度。该平台将提供大约两个订单， 对于类似或更好的测量，测量吞吐量的幅度改进 覆盖率、定量等优于常规的冷凝相液相色谱-质谱联用 LC-MS分析平台，可在数小时内分析数千个样品， 比目前的几周。我们还将调整和完善在我们的生物信息学工具开发 实验室提供自动化和高效的数据处理。最后，该平台将 利用复杂的临床样品在蛋白质组肽/蛋白质覆盖方面进行评价， 测量通量、重复分析的CV和平台稳健性， 到我们实验室现有的领先传统LC-MS平台。

PROJECT SUMMARY/ABSTRACT The objective of this project is to evaluate a new high-throughput and ultra-sensitive platform that will enable more effective and higher throughput proteomics measurements to support and advance cancer research. The new platform will provide substantially higher resolution ion mobility spectrometry (IMS) separations than current platforms. This work also leverages previous developments at our laboratory by combining these new capabilities with more sensitive and efficient ion source and high accuracy MS technologies. The resolution of the IMS achieved will be greatly increased by using electric traveling waves applied to an extremely long path design based upon Structures for Lossless Ion Manipulations (SLIM), while also enabling very high sensitivity to be achieved. The platform will provide approximately two orders of magnitude improvement in measurement throughput for similar or better measurement coverage, quantification, etc. over conventional condensed-phase liquid chromatography-mass spectrometry (LC-MS) platforms, enabling thousands of samples to be analyzed in hours, rather than weeks as at present. We will also adapt and refine bioinformatics tools developed in our laboratory to provide automated and efficient data processing. Finally, the platform will be evaluated utilizing complex clinical samples in terms of proteomic peptide/protein coverage, measurement throughput, CVs of replicate analyses and platform robustness and in comparison to leading-edge conventional LC-MS platforms available in our laboratory.