ANALYTICAL TECHNIQUES FOR CARBOHYDRATE STRUCTURE DETERMINATION

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。 本课程介绍了使用气-液色谱/质谱仪进行糖基残基(组成)和糖基连接(甲基化)分析的技术。参与者对纯化的低聚糖进行了分析，一些参与者带来了他们自己的样本。讲座和演示涵盖了结构分析的其他技术。 此外，还包括使用质谱学或核磁共振光谱学分析糖共轭化合物的模块的选择(见下文描述)。基本的生化技术经验是参加的先决条件。 课程模块 糖结合物的质谱学分析：讨论了利用质谱学对糖结合物进行表征的方法。这一领域包括的主题包括用LC-MS确定占据的糖基化位点，并用顺序的外切糖苷酶消化对糖蛋白多糖进行测序，然后进行MS。就这两个主题进行了实验室演示。 碳水化合物的核磁共振：本模块介绍了多糖和糖共轭化合物的核磁共振波谱的应用，涵盖了i)核磁共振谱的基本特征，ii)用于解析一级结构的方案，iii)当前文献中的应用和例子，以及iv)信息资源。在本模块中，我们使用了四台核磁共振仪器(300 MHz、500 MHz、600 MHz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. This course covered the techniques of glycosyl residue (composition) and glycosyl linkage (methylation) analyses using gas-liquid chromatography/mass spectrometry. Participants performed analyses of purified oligosaccharides, and a few participants brought their own samples. Lectures and demonstrations covered other techniques for structural analysis. In addition, a choice of modules either on using mass spectrometry or NMR spectroscopy for analysis of glycoconjugates were included (see descriptions below). Experience with basic biochemical techniques was a prerequisite for participation. Course Modules Mass Spectrometric (MS) Analysis of Glycoconjugates: The use of mass spectrometry for the characterization of glycoconjugates was discussed. Topics included in this area included identifying occupied glycosylation sites by LC-MS and sequencing glycoprotein glycans with sequential exoglycosidase digestions followed by MS. Laboratory demonstrations were performed on these two topics. NMR of Carbohydrates: This module was an introduction to the application of NMR spectroscopy of 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) were used for demonstrations during this module.