LC-MS Analysis of Site Specific Protein Glycoforms

我们建议优化和传播有针对性的质谱学工作流程，以量化 糖肽和数据解释软件。最新的发展使有效的条带DIA 调整后的裂解条件下的糖肽。这些检测方法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。我们将生成一个知识库 糖肽及其靶向糖肽开发和应用所需的转变 量化工作流。我们将开发必要的软件工具来开发糖肽转换和 分析与这些糖肽转变相关的目标量化工作流程中的光谱。我们会 还制定了用于开发糖肽条带DIA的目标工作流程的分析方案， HR-PRM和LC-MS/MS-MRM分析可供研究界使用。目标客户的可用性 量化资源将首次能够探索疾病背景下的糖蛋白 病理生理学被广泛的研究界所接受。此外，用于定量的质谱分析 位点特异性蛋白糖型的研究将推动糖基化途径和 系统的血糖科学。位点特异性糖蛋白形式的可靠定量将开启靶向研究的新时代 临床化验有可能改变当前的诊断范式并确定新的 在许多糖蛋白靶点中，治疗靶点在很大程度上仍未被探索。

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.