title: An integrated approach using orthogonal analytical techniques to characterize heparan sulfate structure.
abstract: Heparan sulfate (HS), a glycosaminoglycan present on the surface of cells, has been postulated to have important roles in driving both normal and pathological physiologies. The chemical structure and sulfation pattern (domain structure) of HS is believed to determine its biological function, to vary across tissue types, and to be modified in the context of disease. Characterization of HS requires isolation and purification of cell surface HS as a complex mixture. This process may introduce additional chemical modification of the native residues. In this study, we describe an approach towards thorough characterization of bovine kidney heparan sulfate (BKHS) that utilizes a variety of orthogonal analytical techniques (e.g. NMR, IP-RPHPLC, LC-MS). These techniques are applied to characterize this mixture at various levels including composition, fragment level, and overall chain properties. The combination of these techniques in many instances provides orthogonal views into the fine structure of HS, and in other instances provides overlapping / confirmatory information from different perspectives. Specifically, this approach enables quantitative determination of natural and modified saccharide residues in the HS chains, and identifies unusual structures. Analysis of partially digested HS chains allows for a better understanding of the domain structures within this mixture, and yields specific insights into the non-reducing end and reducing end structures of the chains. This approach outlines a useful framework that can be applied to elucidate HS structure and thereby provides means to advance understanding of its biological role and potential involvement in disease progression. In addition, the techniques described here can be applied to characterization of heparin from different sources.
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Daniela
authors: D
authors: Beccati
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Miroslaw
authors: M
authors: Lech
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Jennifer
authors: J
authors: Ozug
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Nur Sibel
authors: NS
authors: Gunay
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Jing
authors: J
authors: Wang
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Elaine Y
authors: EY
authors: Sun
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Joël R
authors: JR
authors: Pradines
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Victor
authors: V
authors: Farutin
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Zachary
authors: Z
authors: Shriver
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA.
authors: Ganesh V
authors: GV
authors: Kaundinya
authors: Momenta Pharmaceuticals Inc., 675 West Kendall Street, Cambridge, MA, 02142, USA. icapila@momentapharma.com.
authors: Ishan
authors: I
authors: Capila
mesh_terms: D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods; D006497:Heparan Sulfate / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods
chemical_list: D006497:Heparan Sulfate
keywords: Analytical; Domains; HPLC; Heparan sulfate; MS; NMR; Structure