Development of a rapid glycosylation profiling system using mass spectrometry dat

摘要： 这项建议的最终目标是开发广泛适用的工具来分析 并将这些工具应用于母乳和母乳的分析 侵袭性的乳腺癌细胞。这项工作将通过完成三个具体的 目标：(1)完成我们启动的全自动糖肽分析工具包 在这笔赠款的第一阶段资金(2)开发大量纳入名单的使用，以 增加MS/MS表征的糖肽离子的数量(3)解决以下两个挑战 当今技术无法实现的糖肽分析：项目1)确定 仅存在于侵袭性乳腺癌的基于糖肽的生物标记物。项目2) 提供母乳中N-和O-连接的糖肽的完整图谱。世界上的大部分 工作将通过结合软件设计、生物样本方面的专业知识来完成 处理、高效液相色谱和质谱学，以开发必要的工具。这些工具的效用 将通过将它们应用于完成与妇女健康相关的项目来展示： 具体地说，我们将对母乳的独特性质提供更详细的见解 比以前提供的更多，我们将表征10-50的糖基化 已知的侵袭性乳腺癌的糖肽标志物。除了我们的贡献之外，我们 为了妇女的健康，这项工作将推动糖蛋白组学领域的发展： 糖蛋白分析方法使生物分析技术能够用于 各种场所，从蛋白质结构/功能分析到药物开发，再到 生物标志物的发现和发展。

Abstract: The ultimate goals of this proposal are to develop broadly applicable tools to analyze glycopeptides in complex matrices and to apply those tools for the analysis of human milk and aggressive breast cancer cells. This work would be accomplished by completing three specific aims: (1) Complete our fully automated glycopeptide analysis tool package, which we started during the first period of this grant's funding (2) Develop the usage of massive inclusion lists to increase the number of glycopeptide ions characterized by MS/MS. (3) Tackle two challenges in glycopeptide analyses that are beyond the reach of today's technology: Project 1) Identify glycopeptide-based biomarkers that are only present in aggressive breast cancer. Project 2) Provide a complete profiling of N- and O-linked glycopeptides in human milk. The bulk of the work would be completed by combining expertise in software design, biological sample handling, HPLC, and mass spectrometry to develop the necessary tools. The utility of the tools would be demonstrated by applying them to complete projects relevant women's health: Specifically, we would provide a more detailed insight into the unique properties of human milk than has ever before been provided, and we would characterize the glycosylation on 10-50 known glycopeptide markers of aggressive breast cancer. In addition to the contributions we make to women's health, this work would drive the field of glycoproteomics forward: The methods for glycoprotein analysis are enabling bioanalytical technologies that can be used in a variety of venues, from protein structure/function analyses to pharmaceutical development, to biomarker discovery and development.