糖胺聚糖：通过液相色谱、毛细管电泳和特异性标记进行寡糖分析

Glycosaminoglycans: oligosaccharide analysis by liquid chromatography, capillary electrophoresis, and specific labeling.

作者信息

Langeslay Derek J, Jones Christopher J, Beni Szabolcs, Larive Cynthia K

机构信息

Department of Chemistry, University of California-Riverside, Riverside, CA, USA.

出版信息

Methods Mol Biol. 2012;836:131-44. doi: 10.1007/978-1-61779-498-8_9.

DOI:10.1007/978-1-61779-498-8_9

PMID:22252632

Abstract

Glycosaminoglycans (GAGs) are a class of biopolymers that include chondrotin sulfate, dermatan sulfate, keratan sulfate, hyaluronic acid, heparin, and heparan sulfate. The GAGs are linear polysaccharides that are microheterogeneous in composition and polydisperse in size. Because they have the most complex structures, this article is aimed at describing a step-by-step procedure for processing and analyzing heparin and heparan sulfate-derived oligosaccharides, although the basic protocols and procedures apply equally well to other members of the GAG family. The methods described in this manuscript include the preparation of oligosaccharides through enzymatic depolymerization, size fractionation by preparative scale size-exclusion chromatography (SEC), and disaccharide isomer analysis by reverse-phase ion-pair high-performance liquid chromatography (RPIP-HPLC) and capillary electrophoresis (CE).

摘要

糖胺聚糖（GAGs）是一类生物聚合物，包括硫酸软骨素、硫酸皮肤素、硫酸角质素、透明质酸、肝素和硫酸乙酰肝素。GAGs是线性多糖，其组成具有微不均一性，大小具有多分散性。由于它们具有最复杂的结构，本文旨在描述一种处理和分析肝素及硫酸乙酰肝素衍生寡糖的分步程序，尽管基本方案和程序同样适用于GAG家族的其他成员。本手稿中描述的方法包括通过酶解聚制备寡糖、通过制备规模尺寸排阻色谱（SEC）进行尺寸分级，以及通过反相离子对高效液相色谱（RPIP-HPLC）和毛细管电泳（CE）进行二糖异构体分析。