SEPARATION AND CHARACTERIZATION OF GLYCOCONJUGATE OLIGOSACCHARIDES

复合糖低聚糖的分离和表征

• 批准号: 7955992
• 负责人: Parastoo Azadi
• 金额: $ 0.13万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7955992
• 关键词: Anions; Area; Biochemical; Biological Assay; Capillary Electrophoresis; Carbohydrates; Chemistry; Chromatography; Computer Retrieval of Information on Scientific Projects Database; Detection; Digestion; Educational process of instructing; Exoglycosidases; Faculty; Funding; Glycoconjugates; Glycolipids; Glycoproteins; Grant; High Pressure Liquid Chromatography; Information Resources; Institutes; Institution; Laboratories; Lectin; Literature; Mass Spectrum Analysis; Monosaccharides; NMR Spectroscopy; Oligosaccharides; Polysaccharides; Protocols documentation; Research; Research Personnel; Resources; Site; Source; Structure; Techniques; Technology; Thin Layer Chromatography; United States National Institutes of Health; Universities; experience; glycosylation; instrument; lectures; sugar

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Separation and Characterization of Glycoconjugate Oligosaccharides (one-week course) This is a one-week course that covers basic techniques for the isolation and characterization of oligosaccharides, including colorimetric quantitation of sugars, chromatographic separation of oligosaccharide mixtures by HPLC, Metrohm peak high-pH anion-exchange chromatography for monosaccharide composition analysis, enzymatic release of oligosaccharides from a glycoconjugate, separation of glycolipids by thin-layer chromatography and detection by overlay assay, and endo- and exoglycosidase treatment of oligosaccharides. The use of lectin blotting techniques for characterization of oligosaccharide structural features are covered. Other topics included are conjugation of oligosaccharides with fluorescent tags, capillary electrophoresis and monosaccharide composition analysis. In addition, modules on using mass spectrometry and/or NMR spectroscopy for analysis of glycoconjugates are included (see descriptions below). Experience with basic biochemical techniques is a prerequisite for participation. In addition to CCRC faculty, other speakers were invited to help teach the course. Dr. Alfred Merrill from Georgia Institute of Technology gave a lecture on "Structural Analysis of Glycolipids by Mass Spectrometry". Dr. Jon Amster from the Chemistry Department at University of Georgia also gave a lecture on "Analysis of oligosaccharides by FT-MS. Additional Course Modules Mass Spectrometric (MS) Analysis of Glycoconjugates: The use of mass spectrometry for the characterization of glycoconjugates is discussed. Topics in this area include identifying occupied glycosylation sites by LC-MS and sequencing glycoprotein glycans with sequential exoglycosidase digestions followed by MS. Laboratory demonstrations are performed on these two topics. NMR of Carbohydrates: The NMR module consists of an introduction to the application of NMR spectroscopy to polysaccharides and glycoconjugates covering (i) essential features of NMR spectra, (ii) protocols used to solve primary structures, (iii) applications and examples from the current literature, and (iv) resources for information. Our four NMR instruments (300-, 500-, 600-, and 800-MHz) are used for demonstrations during this module.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 复合糖寡糖的分离和表征（一周课程） 这是一个为期一周的课程，涵盖寡糖的分离和表征的基本技术， 包括糖的比色定量、通过 HPLC、Metrohm 对寡糖混合物进行色谱分离 用于单糖成分分析、酶促释放的峰值高 pH 阴离子交换色谱 从复合糖中提取寡糖，通过薄层色谱法分离糖脂并通过叠加检测 测定以及寡糖的内切和外切糖苷酶处理。凝集素印迹技术的用途 涵盖了寡糖结构特征的表征。包括的其他主题是结合 带有荧光标签的寡糖、毛细管电泳和单糖成分分析。此外， 包括使用质谱和/或核磁共振波谱分析糖复合物的模块（参见 说明如下）。基本生化技术的经验是参与的先决条件。除了CCRC 教师和其他演讲者被邀请帮助教授该课程。佐治亚理工学院 Alfred Merrill 博士 作了题为“糖脂的质谱结构分析”的讲座。 化学系 Jon Amster 博士 佐治亚大学系也做了题为“Analysis ofoligo Sugars by FT-MS”的讲座。 附加课程模块 糖复合物的质谱 (MS) 分析：利用质谱法表征 讨论了复合糖。该领域的主题包括通过 LC-MS 和测序识别占用的糖基化位点 糖蛋白聚糖，连续外切糖苷酶消化，然后进行 MS。进行实验室演示 关于这两个话题。 碳水化合物的 NMR：NMR 模块包括 NMR 波谱应用的介绍 多糖和糖复合物涵盖 (i) NMR 谱的基本特征，(ii) 用于解决主要问题的方案 结构，(iii) 当前文献中的应用和示例，以及 (iv) 信息资源。我们的四核磁共振 在本模块中，使用仪器（300、500、600 和 800 MHz）进行演示。