Abstract
Capillary electrophoresis is a common technique used for glycosaminoglycan-derived disaccharide analysis because of its high resolving power, high separation efficiency, high sensitivity, short analysis time, and straightforward operation. CE coupled to laser-induced fluorescence (LIF) detection shows an approximately 100 times higher sensitivity than traditional UV detection at 232 nm. 2-Aminoacridone (AMAC) is a widely used fluorophore for labeling unsaturated disaccharides by deductive amination, which is one of the most important method of derivatization of disaccharides for CE-LIF detection. Outlined in this chapter is a protocol of analyzing glycosaminoglycan-derived disaccharides by CE-LIF with AMAC derivatization.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Volpi N, Maccari F, Linhardt RJ (2008) Capillary electrophoresis of complex natural polysaccharides. Electrophoresis 29:3095–3106
Linhardt RJ (1991) Heparin: an important drug enters its seventh decade. Chem Ind 2:45–50
Grag HG, Linhardt RJ, Hales CA (2005) Chemistry and Biology of Heparin and Heparan Sulfate. Elsevier B. V, New York, NY
Linhardt RJ, Turnbull JE, Wang HM, Loganathan D, Gallagher JT (1990) Examination of the substrate specificity of heparin and heparan sulfate lyases. Biochemistry 29:2611–2617
Alicia M, Hitchcock AM, Yates KE, Shortkroff S, Costello CE, Zaia J (2007) Optimized extraction of glycosaminoglycans from normal and osteoarthritic cartilage for glycomics profiling. Glycobiology 17:25–35
Yang B, Solakyildirim K, Chang Y, Linhardt RJ (2011) Hyphenated techniques for the analysis of heparin and heparan sulfate. Anal Bioanal Chem 399:541–557
Rice KG, Kim YS, Grant AC, Merchant ZM, Linhardt RJ (1985) High-performance liquid chromatographic separation of heparin derived oligosaccharides. Anal Biochem 150:325–331
Hileman RE, Smith AE, Toida T, Linhardt RJ (1997) Preparation and structure of heparin lyase-derived heparan sulfate oligosaccharides. Glycobiology 7:231–239
Chuang WL, McAllister H, Rabenstein L (2001) Chromatographic methods for product-profile analysis and isolation of oligosaccharides produced by heparinase-catalyzed depolymerization of heparin. J Chromatogr A 932:65–74
Rice KG, Rottink MK, Linhardt RJ (1987) Fractionation of heparin-derived oligosaccharides by gradient polyacrylamide-gel electrophoresis. Biochem J 244:515–522
Militsopoulou M, Lamari FN, Hjerpe A, Karamanos NK (2002) Determination of twelve heparin- and heparan sulfate-derived disaccharides as 2-aminoacridone derivatives by capillary zone electrophoresis using ultraviolet and laser-induced fluorescence detection. Electrophoresis 23:1104–1109
Mitropoulou TN, Lamari F, Syrokou A, Hjerpe A, Karamanos NK (2001) Identification of oligomeric domains within dermatan sulfate chains using differential enzymic treatments, derivatization with 2-aminoacridone and capillary electrophoresis. Electrophoresis 22:2458–2463
Mao W, Thanawiroon C, Linhardt RJ (2002) Capillary electrophoresis for the analysis of glycosaminoglycans and glycosaminoglycan-derived oligosaccharides. Biomed Chromatogr 16:77–94
Skidmore MA, Guimond SE, Dumax-Vorzet AF, Atrih A, Yates EA, Turnbull JE (2006) High sensitivity separation and detection of heparan sulfate disaccharides. J Chromatogr A 1135:52–56
Lamari FN, Kuhn R, Karamanos NK (2003) Derivatization of carbohydrates for chromatographic, electrophoretic and mass spectrometric structure analysis. J Chromatogr B 793:15–36
Militsopoulou M, Lecomte C, Bayle C, Couderc F, Karamanos NK (2003) Laser-induced fluorescence as a powerful detection tool for capillary electrophoretic analysis of heparin/heparan sulfate disaccharides. Biomed Chromatogr 17:39–41
Mastrogianni O, Lamari F, Syrokou A, Militsopoulou M, Hjerpe A, Karamanos NK (2001) Microemulsion electrokinetic capillary chromatography of sulfated disaccharides derived from glycosaminoglycans. Electrophoresis 22:2743–2745
Hitchcock AM, Bowman MJ, Staples GO, Zaia J (2008) Improved workup for glycosaminoglycan disaccharide analysis using CE with LIF detection. Electrophoresis 29:4538–4548
Zinellu A, Pisanu S, Zinellu E, Lepedda AJ, Cherchi GM, Sotgia S, Carru C, Deiana L, Formato M (2007) A novel LIF-CE method for the separation of hyaluronan- and chondroitin sulfate-derived disaccharides: application to structural and quantitative analyses of human plasma low- and high-charged chondroitin sulfate isomers. Electrophoresis 28:2439–2447
Viola M, Vigetti D, Karousou E, Bartolini B, Genasetti A, Rizzi M, Clerici M, Pallotti F, Luca GD, Passi A (2008) New electrophoretic and chromatographic techniques for analysis of heparin and heparan sulfate. Electrophoresis 29:3168–3174
Yoshida E, Arakawa S, Matsunaga T, Toriumi S, Tokuyama S, Morikawa K, Tahara Y (2002) Cloning, sequencing, and expression of the gene from bacillus circulans that codes for a heparinase that degrades both heparin and heparan sulfate. Biosci Biotechnol Biochem 66:1873–1879
Shaya D, Tocilj A, Li Y, Myette J, Venkataraman G, Sasisekharan R, Cygler M (2006) Crystal structure of heparinase II from Pedobacter heparinus and its complex with a disaccharide product. J Biol Chem 281:15525–15535
Godavarti R, Davis M, Venkataraman G, Cooney C, Langer R, Sasisekharan R (1996) Heparinase III from Flavobacterium heparinum: cloning and recombinant expression in Escherichia coli. Biochem Biophys Res Commun 225:751–758
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Chang, Y., Yang, B., Weyers, A., Linhardt, R.J. (2013). Capillary Electrophoresis for the Analysis of Glycosaminoglycan-Derived Disaccharides. In: Volpi, N., Maccari, F. (eds) Capillary Electrophoresis of Biomolecules. Methods in Molecular Biology, vol 984. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-296-4_7
Download citation
DOI: https://doi.org/10.1007/978-1-62703-296-4_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-295-7
Online ISBN: 978-1-62703-296-4
eBook Packages: Springer Protocols