Shaping MSn Technology for High Impact Glycan Sequencing

塑造高影响力聚糖测序的 MSn 技术

• 批准号: 9753179
• 负责人: VERNON Nye REINHOLD
• 金额: $ 30.07万
• 依托单位: UNIVERSITY OF NEW HAMPSHIRE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-18 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753179
• 关键词: Academia; Anabolism; Antibodies; Area; Binding; Biological; Biological Models; Biological Products; Biomedical Research; Carbohydrates; Cell physiology; Chemistry; Collection; Communities; Data; Development; Diffusion; Disease; Engineering; Epitopes; Fucose; Glycobiology; Glycoproteins; Glycosaminoglycans; Glycoside Hydrolases; Glycosphingolipids; Heparin; Industry; Internet; Ions; Laboratories; Learning; Lectin; Libraries; Link; Mass Spectrum Analysis; Metadata; Methylation; Milk; Oligosaccharides; Parasites; Play; Polymers; Polysaccharides; Positioning Attribute; Process; Protocols documentation; Publishing; Reporting; Role; Sampling; Scheme; Scientist; Series; Shapes; Sialic Acids; Software Tools; Source; Structure; System; Technology; Time; Unspecified or Sulfate Ion Sulfates; Work; analog; bacterial H antigen; base; biological research; biomarker discovery; cancer biomarkers; carbohydrate structure; computer code; computerized data processing; data acquisition; data sharing; design; glycosylation; host-microbe interactions; insight; instrument; lactosamine; software systems; stable isotope; synergism; technology diffusion; tool; vaccine discovery; web portal; web-enabled

Project Summary/Abstract Ion Trap Sequential Mass Spectrometry (IT-MSn) based sequencing has been proven a powerful technology for detailed carbohydrate structural characterization. Different from many traditional oligosaccharide analysis strategies, MSn sequencing does not depend on glycan biosynthesis rules, or inferential data. This unique feature is particularly beneficial in studying abnormal glycosylation in diseases, or engineering glycotherapeutics glycosylation machinery in non-conventional expression systems, because glycosylation in these systems are less studied, biosynthesis rules established from model systems may not be applicable any more. The appreciation of the value provided by MSn sequencing keeps growing in both academia and industry. Nevertheless, the steep learning curve of MSn sequencing has been hindering the wide adaptation of the technology. This laboratory has pioneered this glycan sequencing technology and is uniquely positioned to advance and disseminate this approach to the glycobiology community. In an attempt to flatten the learning curve, we propose four Specific Aims in the application: (A) to expand MSn library collection and build a web portal to enable web access; (B) to develop an integrated software system for automating MSn glyco-epitope identification; (C) to seek synergy with traditional oligosaccharide structural analysis strategies; (D) to extend detailed MSn sequencing to cover glycosaminoglycan (GAG) related polymers. The deliverables of the proposed projects will be a set of software tools and established lab protocols that enable non-specialized biomedical labs to conduct detailed carbohydrate characterization using MSn sequencing. We believe the proposed developments are in line with the RFA-RM-16-023, and will accelerate the diffusion of the technology.

项目摘要/摘要 基于离子陷阱序列质谱仪(IT-MSN)的测序已经被证明是一种 用于详细描述碳水化合物结构的强大技术。与许多不同的是 传统的寡糖分析策略，MSN测序不依赖于多糖 生物合成规则，或推论数据。这一独特的特点在学习中特别有益 疾病中的糖基化异常，或工程糖疗法中的糖基化机制 非常规表达系统，因为这些系统中的糖基化研究较少， 从模型系统建立的生物合成规则可能不再适用。这个 对MSN测序提供的价值的认识在学术界和 工业。然而，MSN测序的陡峭学习曲线一直在阻碍 该技术具有广泛的适应性。这个实验室开创了糖链测序的先河 技术，并处于独特的地位来推动和传播这种方法，以 糖生物学社区。为了使学习曲线变得平坦，我们提出了四个具体的建议 应用程序的目标：(A)扩大MSN图书馆馆藏并建立门户网站，以实现 网络接入；(B)开发一个自动化MSN糖基表位的综合软件系统 (C)寻求与传统低聚糖结构分析策略的协同作用； (D)扩大MSN的详细测序范围，以涵盖与糖胺聚糖(GAG)相关的聚合物。 拟议项目的可交付成果将是一套软件工具和已建立的实验室 使非专业生物医学实验室能够进行详细的碳水化合物 使用MSN测序进行表征。我们相信建议的发展项目符合 RFA-RM-16-023，并将加速该技术的传播。