LC-MS Analysis of Site Specific Protein Glycoforms

位点特异性蛋白质糖型的 LC-MS 分析

• 批准号: 9761502
• 负责人: NATHAN J EDWARDS
• 金额: $ 49.46万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-09 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9761502
• 关键词: Adoption; Biological; Biological Assay; Biomedical Research; Calibration; Cell Line; Cells; Clinical; Communities; Complex; Computer software; Data Analyses; Development; Diagnostic; Disease; Engineering; Functional disorder; Generations; Glycoconjugates; Glycopeptides; Glycoproteins; Heterogeneity; Human; Ions; Knowledge; Laboratories; Libraries; Liver Cirrhosis; Methods; Modification; Mucins; Organism; Outcome; Pathway interactions; Post-Translational Protein Processing; Process; Protein Glycosylation; Proteins; Protocols documentation; Publishing; Research; Research Personnel; Resources; Sampling; Sensitivity and Specificity; Serum; Site; Software Tools; Specificity; Standardization; System; Time; Tissues; cell type; clinically relevant; design; glycoproteomics; glycosylation; improved; innovation; instrument; interest; knowledge base; new therapeutic target; non-invasive monitor; novel; protocol development; software development; tool

We propose to optimize and disseminate targeted mass spectrometric workflows for the quantification of glycopeptides and software for data interpretation. Recent developments enable efficient SWATH DIA of glycopeptides under adjusted fragmentation conditions. The sensitivity of these assays can be further improved in targeted LC-MS/MS-MRM and HR-PRM workflows. We will generate a knowledgebase of glycopeptides and the transitions needed for the development and application of the targeted glycopeptide quantification workflows. We will generate software tools necessary to develop glycopeptide transitions and analyze spectra from targeted quantification workflows with respect to these glycopeptide transitions. We will also make the analytical protocols for the development of targeted workflows for glycopeptide SWATH DIA, HR-PRM, and LC-MS/MS-MRM assays available to the research community. The availability of targeted quantification resources will enable, for the first time, exploration of glycoproteins in the context of disease pathophysiology by the broad research community. Furthermore, mass spectrometric assays for quantification of site specific protein glycoforms will drive research in the characterization of glycosylation pathways and systems glycoscience. Reliable quantification of site-specific glycoproteoforms will begin a new era of targeted clinical assays that have the potential to change the current diagnostic paradigm and to identify new therapeutic targets among the many glycoprotein targets that remain largely unexplored.

我们建议优化和传播靶向质谱工作流程，以量化 糖肽和数据解释软件。最近的发展使有效的小水线面双体船直径 在调整的片段化条件下的糖肽。这些测定的灵敏度可以进一步提高。 改进了目标LC-MS/MS-MRM和HR-PRM工作流程。我们将建立一个知识库， 糖肽及其开发和应用所需的转换 量化工作流。我们将生成开发糖肽转换所需的软件工具， 根据这些糖肽转换分析来自目标定量工作流程的光谱。我们将 还为糖肽SWATH DIA的靶向工作流程的开发制定分析方案， HR-PRM和LC-MS/MS-MRM分析可供研究界使用。有针对性的可用性 量化资源将首次在疾病背景下探索糖蛋白 病理生理学被广泛的研究界。此外，用于定量的质谱测定 位点特异性蛋白糖型的研究将推动糖基化途径的表征研究， 系统糖科学位点特异性糖蛋白的可靠定量将开始一个新的靶向 具有改变当前诊断模式和鉴定新的 在许多糖蛋白靶点中的治疗靶点仍然在很大程度上未被探索。