High Resolution High Throughput Proteomics Platform for Cancer Research

项目摘要/摘要 该项目的目标是评估一种新的高通量和超灵敏平台 这将使蛋白质组学测量能够更有效和更高吞吐量地支持和 推进癌症研究。新的平台将提供更高的分辨率 移动光谱(IMS)的分离比目前的平台更好。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MS技术。IMS的分辨率 通过使用电行波应用到极长的距离将大大提高实现 基于无损离子操纵结构的路径设计(SLIM)，同时还实现 要达到非常高的灵敏度。该平台将提供大约两个订单 类似或更好的测量的测量吞吐量的幅度提高 常规凝聚相液相色谱质谱的覆盖率、定量等 光谱(LC-MS)平台，使数千个样品能够在几个小时内进行分析，而不是 比目前的几个星期要多。我们还将调整和改进在我们的 实验室提供自动化和高效的数据处理。最后，平台将是 利用复杂的临床样本在蛋白质组肽/蛋白质覆盖率方面进行评估， 测量吞吐量、复制分析的CV和平台稳健性并进行比较 到我们实验室提供的尖端传统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.