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Multiplex Mass Spectrometric Protein Assays for Precise Monitoring of the Pathophysiology of Obesity

Multiplex Mass Spectrometric Protein Assays for Precise Monitoring of the Pathophysiology of Obesity
用于精确监测肥胖病理生理学的多重质谱蛋白质分析
批准号:
10020391
负责人:
Wei-Jun Qian
金额:
$75.56万
依托单位国家:
美国
项目类别:
财政年份:
2019
资助国家:
美国
项目状态:
已结题
起止时间:
2019-09-20 至 2023-07-31

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中文摘要
翻译
项目概要/摘要 近几十年来,儿童和青少年超重和肥胖的患病率急剧增加, 成年人,达到流行病的比例。为了对肥胖的病理生理学有新的认识 为了更好的个性化患者护理,迫切需要开发可靠的多重蛋白质测定, 可以很容易地在临床实验室中实施,以实现对许多激素的精确监测, 与肥胖密切相关的炎症标志物。不幸的是,目前的临床测定通常缺乏 重复性,特别是由于缺乏测定标准化而在不同的临床实验室之间。 靶向质谱法是一种有前途的技术,可为血液蛋白质提供稳健的多重测定。 因此,本申请的总体目标是开发和验证可重现和可转移的质量 基于光谱的多重蛋白质测定,用于精确定量一组肥胖标志物。 该小组将涵盖肥胖病理生理学的多个方面,包括葡萄糖稳态和能量 平衡(例如,胰岛素、瘦素和其它激素),促炎标记物(例如,C-反应蛋白),以及 抗炎标记物(例如,脂联素)。为了便于全面验证稳健性和可转移性, 在这些测定中,将进行实验室间测定验证。具体而言,目标1将侧重于初始检测 开发最佳配置,并证明内源性 血浆中的分析物。目标2将集中在分析优化和完整的分析表征, 重现性、稳定性、选择性、线性和定量限方面。目标3将展示 通过实验室间定量内源性分析物在临床血浆试验队列中的应用 来自正常体重和肥胖受试者以及体重减轻前后的肥胖个体的样品, 针对选定的分析物如胰岛素和瘦素的成熟的免疫测定进行基准测试。到 便于在其他实验室轻松实施经验证的测定,所有测定数据包括 色谱数据和详细的标准操作程序将通过公共数据共享 储存库和分析门户。总之,该项目将建立强大的和易于转移的多重测定 这将使肥胖症的病理生理学的精确监测。
英文摘要
PROJECT SUMMARY/ABSTRACT The prevalence of overweight and obesity has increased dramatically in recent decades in both children and adults, reaching an epidemic proportion. In order to gain new insights into the pathophysiology of obesity towards better personalized patient care, there is a critical need to develop reliable multiplex protein assays that can be easily implemented in clinical laboratories to enable precise monitoring of many hormones and inflammatory markers closely associated with obesity. Unfortunately, current clinical assays often lack of reproducibility, especially between different clinical laboratories due to the lack of assay standardization. Targeted mass spectrometry is a promising technology for providing robust multiplex assays for blood proteins. Therefore, the overall objective of this application is to develop and validate reproducible and transferable mass spectrometry-based multiplex protein assays for enabling precise quantification of a panel of obesity markers. The panel will cover multiple aspects of obesity pathophysiology, including glucose homeostasis and energy balance (e.g., insulin, leptin, and other hormones), pro-inflammatory markers (e.g., C-reactive protein), and anti-inflammatory markers (e.g., adiponectin). To facilitate full validation of the robustness and transferability of the assays, an inter-lab assay validation will be pursued. Specifically, Aim 1 will be focused on initial assay development for optimal configurations and on demonstrating confident multiplex detection of endogenous analytes in blood plasma. Aim 2 will be centered on assay optimization and full assay characterization in the aspects of reproducibility, stability, selectivity, linearity, and limit of quantification. Aim 3 will demonstrate the utility of the assays through inter-lab quantification of endogenous analytes in a pilot cohort of clinical plasma samples from normal weight and obese subjects, and obese individual before and after weight loss and benchmarking against well-established immunoassays for selected analytes such as insulin and leptin. To facilitate the easy- implementation of the validated assays in other laboratories, all assay data including chromatographic data and detailed standard operating procedures will be shared through public data repositories and assay portals. Together, the project will establish robust and easy-to-transfer multiplex assays that will enable precise monitoring of the pathophysiology of obesity.
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