Faculty and Student Publications
Document Type
Article
Publication Date
9-1-2019
Abstract
© 2019, The Author(s). Heparin and heparan sulfate (Hp/HS) are linear complex glycosaminoglycans which are involved in diverse biological processes. The structural complexity brings difficulties in separation, making the study of structure-function relationships challenging. Here we present a separation method for Hp/HS oligosaccharide fractionation with cross-compatible solvent and conditions, combining size exclusion chromatography (SEC), ion-pair reversed phase chromatography (IPRP), and hydrophilic interaction chromatography (HILIC) as three orthogonal separation methods that do not require desalting or extensive sample handling. With this method, the final eluent is suitable for structure-function relationship studies, including tandem mass spectrometry and microarray printing. Our data indicate that high resolution is achieved on both IPRP and HILIC for Hp/HS isomers. In addition, the fractions co-eluted in IPRP could be further separated by HILIC, with both separation dimensions capable of resolving some isomeric oligosaccharides. We demonstrate this method using both unpurified reaction products from isomeric synthetic hexasaccharides and an octasaccharide fraction from enoxaparin, identifying isomers resolved by this multi-dimensional separation method. We demonstrate both structural analysis by MS, as well as functional analysis by microarray printing and screening using a prototypical Hp/HS binding protein: basic-fibroblast growth factor (FGF2). Collectively, this method provides a strategy for efficient Hp/HS structure-function characterization.
Relational Format
journal article
Recommended Citation
Ilhan, M., Ali, Z., Khan, I. A., Taştan, H., & Küpeli Akkol, E. (2019). Promising activity of Anthemis austriaca Jacq. on the endometriosis rat model and isolation of its active constituents. Saudi Pharmaceutical Journal, 27(6), 889–899. https://doi.org/10.1016/j.jsps.2019.06.002
DOI
10.1016/j.jsps.2019.06.002