SEPARATION AND CHARACTERIZATION OF GLYCOCONJUGATE OLIGOSACCHARIDES

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 糖共轭低聚糖的分离与表征(一周课程) 这是一个为期一周的课程，涵盖了低聚糖分离和表征的基本技术， 包括糖的比色定量，低聚糖混合物的高效液相色谱分离，Metrohm 峰高pH阴离子交换层析分析单糖组成，酶促释放 糖偶联物中的低聚糖，糖脂的薄层分离和重叠检测 低聚糖的内切和外切糖苷酶处理。凝集素印迹技术的应用 低聚糖的结构特征的表征也包括在内。包括的其他主题是 低聚糖的荧光标记，毛细管电泳法和单糖组成分析。此外, 包括使用质谱学和/或核磁共振光谱学分析糖共轭化合物的模块(请参阅 描述如下)。基本的生化技术经验是参加的先决条件。除了CCRC之外 教职员工和其他演讲者被邀请帮助讲授这门课程。佐治亚理工学院的Alfred Merrill博士 主讲《糖脂的质谱学结构分析》。化学学会的乔恩·阿姆斯特博士 佐治亚大学的系也作了题为《FT-MS分析低聚糖》的讲座。 附加课程模块 糖偶联物的质谱学分析：使用质谱学来表征糖偶联物 对糖共轭化合物进行了讨论。这一领域的主题包括用LC-MS鉴定占据的糖基化位点和测序 糖蛋白多糖与外切糖苷酶顺序消化，然后MS实验室演示 在这两个主题上。 碳水化合物的核磁共振：核磁共振模块包括对核磁共振波谱应用于 多糖和糖结合物(I)核磁共振谱的基本特征，(II)用于解决初级问题的方案 结构；(3)现有文献中的应用和例子；(4)信息资源。我们的四个核磁共振 在本模块中使用仪器(300-、500-、600-和800-MHz)进行演示。

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.