LC-IMS-MS Techniques for Human Plasma Profiling

用于人体血浆分析的 LC-IMS-MS 技术

• 批准号: 6989473
• 负责人: Stephen Valentine
• 金额: $ 13.69万
• 依托单位: PREDICTIVE PHYSIOLOGY AND MEDICINE, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2006-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6989473
• 关键词: biomarker; cardiovascular disorder; clinical research; high throughput technology; human subject; mass spectrometry; plasma; protein quantitation /detection; proteomics; technology /technique development

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.

描述（由申请人提供）：将开发一种新的高通量、高覆盖率蛋白质组技术，利用离子淌度/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 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，该图将提供更高的等离子体成分分辨率（10 到 50 倍）。工作重点是确定群体中不同个体以及特定个体随测量次数 (N) 变化的蛋白质丰度随时间变化的变化。新技术缩短了实验时间，使多次测量成为可能。随着 N 的增加，蛋白质分配和丰度测定的置信度也会增加。此外，执行必要的氮测量来定义“正常”的能力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），因为将更准确地了解被认为是统计相关的变化。完成第一阶段研究中提出的工作有三个具体目标。这些是： 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现性并细化分配； 3) 开始比较“正常”和“患病”图谱，以描绘新的生物标志物（主要在 II 期研究中进行）。构建更高分辨率的高覆盖率蛋白质组图谱将允许更深入、更广泛地了解血浆蛋白质组。这样的有利位置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以下方面提供线索： 与心血管疾病相关的生物标志物，可能与诊断、治疗监测以及药物发现研究相关。