Microscale separation of heparosan, heparan sulfate, and heparin

Abstract

The separation and quantification of glycosaminoglycan (GAG) chains with different levels of sulfation from cells and media, and prepared through chemoenzymatic synthesis or metabolic engineering, pose a major challenge in glycomics analysis. A method for microscale separation and quantification of heparin, heparan sulfate, and heparosan from cells is reported. This separation relies on a mini strong anion exchange spin column eluted stepwise with various concentrations of sodium chloride. Disaccharide analysis by LC-MS was used to monitor the chemical structure of the various GAG chains that were recovered.