Cell-Based Microscale Isolation of Glycoaminoglycans for Glycomics Study

J Carbohydr Chem. 2012 May;31(4-6):420-435. doi: 10.1080/07328303.2012.658126.

Abstract

Glycomics research requires the isolation of glycans from cells for structural characterization and functional studies of the glycans. A method for cell-based microscale isolation and quantification of highly sulfated, moderately sulfated, and nonsulfated glycosaminoglycans (GAGs) was developed using Chinese hamster ovary (CHO) cells. This microscale isolation relies on a mini-strong anion exchange spin column eluted stepwise with different concentrations of sodium chloride solution. Hyaluronic acid, chondroitin sulfate, and heparin were used to optimize the isolation of the endogenous glycosaminoglycans in CHO cells. This method can also be used to determine the presence of nonsulfated GAGs including heparosan, hyaluronic acid, and nonsulfated chondroitin.

Keywords: Anion exchange chromatography; Chinese hamster ovary cells; Disaccharide analysis; Glycosaminoglycans; Heparosan.