ANALYTICAL TECHNIQUES FOR CARBOHYDRATE STRUCTURE DETERMINATION

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH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 中心，不一定是研究者的机构。 本课程涵盖了糖基残基（组成）和糖基连接（甲基化）的分析技术，使用气液色谱/质谱。 参与者对纯化的低聚糖进行了分析，一些参与者带来了自己的样品。 讲座和演示涵盖了结构分析的其他技术。 此外，还包括使用质谱法或NMR光谱法分析糖缀合物的模块选择（见下文描述）。 基本生物化学技术的经验是参加的先决条件。 课程模块 糖缀合物的质谱（MS）分析：讨论了质谱法用于糖缀合物表征的用途。 这一领域的主题包括通过LC-MS确定已占据的糖基化位点，并通过连续的外切糖苷酶diglase和MS对糖蛋白聚糖进行测序。对这两个主题进行了实验室演示。 碳水化合物的NMR：该模块介绍了多糖和糖缀合物的NMR光谱应用，涵盖i）NMR光谱的基本特征，ii）用于解决一级结构的协议，iii）当前文献中的应用和示例，以及iv）信息资源。 我们的四台NMR仪器（300-，500-，600-和800 MHz）在本模块中用于演示。

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.