Skidmore Mark, Atrih Abdel, Yates Ed, Turnbull Jeremy E
School of Biological Sciences, University of Liverpool, Liverpool, UK.
Methods Mol Biol. 2009;534:157-69. doi: 10.1007/978-1-59745-022-5_12.
The analysis of heparin/HS saccharides derived from small amounts of tissue or cells is a considerable technical challenge and the development of methods to characterise these carbohydrates has progressed comparatively slowly. A number of procedures have been devised to tag glycans selectively at the reducing end with a group that will enhance the sensitivity of detection and facilitate chromatographic separations. Outlined in this chapter are two useful strategies designed specifically for the analysis of heparin/HS saccharides. The first involves a fluorophore label, Bodipy-FL-hydrazide, which permits highly sensitive (fmol level) detection of saccharides utilising high performance strong anion exchange chromatography. The second facilitates oligosaccharide separation by gel-permeation chromatography and reverse phase high performance ion-pairing chromatography (RP-HPIPC) through the use of a phenylsemicarbazide tag. The latter also serves as an effective mass tag for electrospray mass spectrometry, permitting enhanced analysis of HS saccharides. These methods provide new opportunities for the development of glycomics approaches to study the structure and function of the heparan sulfate family of glycans.
对源自少量组织或细胞的肝素/硫酸乙酰肝素糖进行分析是一项颇具技术挑战性的工作，用于表征这些碳水化合物的方法发展相对缓慢。人们已设计出多种程序，以便在还原端用一个能提高检测灵敏度并便于色谱分离的基团对聚糖进行选择性标记。本章概述了专门为肝素/硫酸乙酰肝素糖分析设计的两种有用策略。第一种策略涉及一种荧光团标记物，即Bodipy-FL-酰肼，它可利用高效强阴离子交换色谱对糖类进行高灵敏度（飞摩尔水平）检测。第二种策略通过使用苯氨基脲标签，借助凝胶渗透色谱和反相高效离子对色谱（RP-HPIPC）来促进寡糖分离。后者还可作为电喷雾质谱的有效质量标签，从而增强对硫酸乙酰肝素糖的分析。这些方法为开发糖组学方法以研究硫酸乙酰肝素聚糖家族的结构和功能提供了新机遇。
