Cell-Based Microscale Isolation of Glycoaminoglycans for Glycomics Study.
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.
College of Food Science and Technology, Ocean University of China, Qingdao, Shandong, 266003, P.R. China ; Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Xue
X
Zhao
Bo
B
Yang
Payel
P
Datta
Leyla
L
Gasmili
Fuming
F
Zhang
Robert J
RJ
Linhardt
Anion exchange chromatography; Chinese hamster ovary cells; Disaccharide analysis; Glycosaminoglycans; Heparosan