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（IMS-MS）来表征血浆蛋白质组。IMS-MS仪器将用于通过创建通过分析200份血浆样本获得的高维分析图谱来解决血浆蛋白质组的复杂性。与商业技术相比，该图将提供更高的血浆成分分辨率（10至50倍）。工作将集中在确定蛋白质丰度的变异性，作为测量数量（N）的函数，为整个人口中的不同个体，以及为特定的个人作为时间的函数。通过新技术提供的减少的实验时间尺度，多个测量成为可能。随着N的增加，蛋白质分配和丰度测定的置信水平也增加。此外，进行必要的N测量以定义“正常”的能力将有助于确定心血管疾病样品中的蛋白质丰度变化（II期研究），因为将更准确地知道什么被认为是统计学相关的变化。完成第一阶段研究建议的工作有三个具体目标。这些措施是：1）开发LC-IMS-（CID）n-TOF仪器作为高通量血浆蛋白质组分析的稳健平台; 2）评估“正常”血浆蛋白质组图谱的再现性和精确分配;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II期研究中进行）。构建高覆盖率、高分辨率的蛋白质组图谱，将使人们对血浆蛋白质组有更深、更广的了解。这样的Vantage对于确定心血管疾病样本中蛋白质丰度的变化至关重要（II期研究）。观察到的变化可能为心血管疾病相关的生物标志物提供线索，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.