LC-IMS-MS Techniques for Human Plasma Profiling

描述(由申请人提供)：将开发一种新的高通量、高覆盖率的蛋白质组学技术，该技术利用离子迁移率/质谱学(IMS-MS)来表征血浆蛋白质组。IMS-MS仪器将通过创建通过分析200个血浆样本获得的高维分析图来解决血浆蛋白质组的复杂性。与商业技术相比，这张地图将提供更高的等离子体成分分辨率(分辨率为10到50倍)。工作将集中在确定蛋白质丰度的变异性作为种群中不同个体的测量次数(N)的函数，以及特定个体作为时间的函数。这项新技术减少了实验时间刻度，使多次测量成为可能。随着氮的增加，蛋白质分配和丰度测定的置信度也增加。此外，能够进行必要的N测量来定义“正常”将有助于确定心血管疾病样本中蛋白质丰度的变化(第二阶段研究)，因为被认为是统计学上相关的变化将被更准确地知道。完成第一阶段研究中提出的工作有三个具体目标。它们是：1)开发LC-IMS-(CID)n-TOF仪器作为高通量血浆蛋白质组分析的强大平台；2)评估“正常”血浆蛋白质组图谱的重复性并改进其赋值；以及3)启动“正常”和“患病”图谱的比较，以描绘新的生物标志物(主要在第二阶段研究中进行)。构建更高分辨率的高覆盖率蛋白质组图谱将使人们能够更深入、更广泛地了解血浆蛋白质组。这样的有利条件对于确定心血管疾病样本中蛋白质丰度的变化至关重要(第二阶段研究)。观察到的变化可能提供与心血管疾病相关的生物标记物的线索，并可能与诊断、治疗监测以及药物发现研究相关。

DESCRIPTION (provided by applicant): A New high-throughput, high-coverage proteomics technology that utilizes ion mobility/mass spectrometry (IMS-MS) for the characterization of the plasma proteome will be developed. The IMS-MS instrumentation will be utilized to address the complexity of the plasma proteome by creating a high-dimension, analytical map obtained by analysis of 200 plasma samples. This map will offer a higher resolution of plasma components when compared with commercial technology (by factors of 10 to 50). Work will center on determining the variability in protein abundances as a function of the number of measurements (N) for different individuals across a population as well as for specific individuals as a function of time. The multiple measurements are made possible by a decreased experimental timescale afforded by the new technology. As N increases, the confidence level of protein assignments and abundance determinations also increases. Additionally the ability to perform the requisite N measurements to define "normal" will facilitate the determination of protein abundance change in cardiovascular disease samples (Phase II studies) as what is considered a statistically relevant change will be more accurately known. There are three specific aims to accomplish the work proposed in Phase I studies. These are: 1) Develop LC-IMS-(CID)n-TOF instrumentation as a robust platform for high-throughput plasma proteome analysis; 2) Assess the reproducibility and refine assignments of the "normal" plasma proteome map; and, 3) Initiate comparison of normal" and "diseased" maps for delineation of new biomarkers (primarily carried out in Phase II studies). The construction of a high-coverage proteome map of higher resolution will allow a deeper, broader view of the plasma proteome. Such a vantage point is vital for the determination of protein abundance changes in cardiovascular disease samples (Phase II studies). Observed changes may provide clues to biomarkers associated with cardiovascular disease and may have relevance for diagnostics, therapeutic monitoring, as well as drug discovery studies.