MS based approaches for diabetes biomarker discovery

基于 MS 的糖尿病生物标志物发现方法

• 批准号: 7617578
• 负责人: Urban A Kiernan
• 金额: $ 26.19万
• 依托单位: INTRINSIC BIOPROBES, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2010-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7617578
• 关键词: Affinity; Biological; Biological Assay; Biological Markers; Bioprobe; Characteristics; Clinical; Complex; Data; Detection; Diabetes Mellitus; Digestion; Event; Figs - dietary; Frequencies; Goals; Individual; Ligands; Link; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Metals; Methodology; Methods; Monoclonal Antibodies; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Insulin-Dependent Diabetes Mellitus; OGTT; Phase; Plasma Cells; Post-Translational Protein Processing; Prediabetes syndrome; Process; Proteins; Proteomics; Relative (related person); Request for Applications; Research; Robotics; Sampling; Screening procedure; Solutions; Specificity; Statistical Methods; Study Section; Surface; Technology; Validation; Variant; base; clinical application; cohort; diabetic; disease diagnosis; novel; programs; protein profiling; validation studies

DESCRIPTION (provided by applicant): The objective of this proposal is to apply novel affinity mass spectrometry (AMS) technologies to identify protein biomarkers for pre-diabetes or type 2 diabetes. The methodology used combines micro-affinity protein capture from complex biological solutions with mass spectrometric detection. In this first phase (R21), we will screen the provided plasma and blood cell samples to identify candidate proteins, and protein profiles, that can appropriately differentiate sample origin from 1 of the 3 groups (normal, pre-diabetic and diabetic; as determined by the administration of an oral glucose tolerance test). Both plasma and blood cell samples will be assayed with affinity pipettes derivatized with poly- and monoclonal antibodies for specific protein target analysis. Additionally, groups of proteins will be analyzed via hydrophilic and hydrophobic ligands, metal chelating ligands, and other wide-specificity affinity surfaces. Each of these ligand surfaces possesses specific affinity characteristics that will result in the capture and purification of individual or whole groups of proteins. Protein variations observed as a result of qualitative and/or quantitative differences, that have statistical significance and correctly group samples according to their origin, will be viewed as potential biomarkers. In the second (validation) phase (R33), we will validate the discovered biomarker(s) via the application of the same methods and approaches to a larger group of sample cohorts through the use of high throughput parallel robotic processing. In the event that multiple protein biomarkers are validated, AMS panels for diabetes will be developed. This will be achieved through the construction of multi-analyte affinity pipettes that will selectively retrieve the targeted biomarkers that differentiate healthy from diseased states. Such panels will also be subject to the same validation process that each of its components endured. The ultimate result of this research will be validated protein biomarkers for pre-and type 2 diabetes and technology that are ready for clinical screening and diagnosis of the disease.

描述（由申请人提供）：本提案的目的是应用新型亲和质谱（AMS）技术来识别糖尿病前期或 2 型糖尿病的蛋白质生物标志物。所使用的方法将复杂生物溶液中的微亲和蛋白捕获与质谱检测相结合。在第一阶段 (R21) 中，我们将筛选提供的血浆和血细胞样本，以识别候选蛋白质和蛋白质谱，这些蛋白质和蛋白质谱可以适当地区分 3 组（正常、糖尿病前期和糖尿病；根据确定）中的一组的样品来源。通过进行口服葡萄糖耐量试验）。血浆和血细胞样品都将使用用多克隆和单克隆抗体衍生的亲和移液器进行分析，以进行特定的蛋白质目标分析。此外，还将通过亲水性和疏水性配体、金属螯合配体和其他广泛特异性的亲和表面来分析蛋白质组。这些配体表面中的每一个都具有特定的亲和特性，这将导致捕获和纯化单个或整个蛋白质组。由于定性和/或定量差异而观察到的蛋白质变异，具有统计显着性并根据其来源对样品进行正确分组，将被视为潜在的生物标志物。在第二（验证）阶段 (R33)，我们将通过使用高通量并行机器人处理对更大的样本组应用相同的方法和方法来验证发现的生物标志物。如果多种蛋白质生物标志物得到验证，将开发用于糖尿病的 AMS 组合。这将通过构建多分析物亲和移液器来实现，该移液器将选择性地检索区分健康状态和疾病状态的目标生物标志物。此类面板还将接受其每个组件所经历的相同验证过程。这项研究的最终结果将是针对 2 型糖尿病前期和 2 型糖尿病的经过验证的蛋白质生物标志物以及为该疾病的临床筛查和诊断做好准备的技术。

项目成果

Quantitation of target proteins and post-translational modifications in affinity-based proteomics approaches.

基于亲和力的蛋白质组学方法中目标蛋白的定量和翻译后修饰。

• 发表时间: 2007-06
• 作者: Kiernan; Urban A
• 通讯作者: Urban A