A chip-based amide-HILIC LC/MS platform for glycosaminoglycan glycomics profiling.
A key challenge to investigations into the functional roles of glycosaminoglycans (GAGs) in biological systems is the difficulty in achieving sensitive, stable, and reproducible mass spectrometric analysis. GAGs are linear carbohydrates with domains that vary in the extent of sulfation, acetylation, and uronic acid epimerization. It is of particular importance to determine spatial and temporal variations of GAG domain structures in biological tissues. In order to analyze GAGs from tissue, it is useful to couple MS with an on-line separation system. The purposes of the separation system are both to remove components that inhibit GAG ionization and to enable the analysis of very complex mixtures. This contribution presents amide-silica hydrophilic interaction chromatography (HILIC) in a chip-based format for LC/MS of heparin, heparan sulfate (HS) GAGs. The chip interface yields robust performance in the negative ion mode that is essential for GAGs and other acidic glycan classes while the built-in trapping cartridge reduces background from the biological tissue matrix. The HILIC chromatographic separation is based on a combination of the glycan chain lengths and the numbers of hydrophobic acetate (Ac) groups and acidic sulfate groups. In summary, chip based amide-HILIC LC/MS is an enabling technology for GAG glycomics profiling.
Staples
Gregory O
GO
Department of Biochemistry, Boston University School of Medicine, Mass Spectrometry Resource, Boston, MA 02118, USA.
Bowman
Michael J
MJ
Costello
Catherine E
CE
Hitchcock
Alicia M
AM
Lau
James M
JM
Leymarie
Nancy
N
Miller
Christine
C
Naimy
Hicham
H
Shi
Xiaofeng
X
Zaia
Joseph
J
D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D054794:Glycomics / Q000379:methods*; D006025:Glycosaminoglycans / Q000032:analysis*; D006497:Heparan Sulfate / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013552:Swine
D006025:Glycosaminoglycans; D006497:Heparan Sulfate