FACE Analysis as a Fast and Reliable Methodology to Monitor the Sulfation and Total Amount of Chondroitin Sulfate in Biological Samples of Clinical Importance
by
Evgenia Karousou 1,*, Athanasia Asimakopoulou 2, Luca Monti 3, Vassiliki Zafeiropoulou 2, Nikos Afratis 2, Panagiotis Gartaganis 2, Antonio Rossi 3, Alberto Passi 1,* and Nikos K. Karamanos 2,*
Evgenia Karousou 1,*, Athanasia Asimakopoulou 2, Luca Monti 3, Vassiliki Zafeiropoulou 2, Nikos Afratis 2, Panagiotis Gartaganis 2, Antonio Rossi 3, Alberto Passi 1,* and Nikos K. Karamanos 2,*
1
Department of Surgical and Morphological Sciences, University of Insubria, 21100 Varese, Italy
2
Laboratory of Biochemistry, Department of Chemistry, University of Patras, 26504 Patras, Greece
3
Department of Molecular Medicine, Unit of Biochemistry, University of Pavia, 27100 Pavia, Italy
*
Authors to whom correspondence should be addressed.
Molecules 2014, 19(6), 7959-7980; https://doi.org/10.3390/molecules19067959
Received: 15 April 2014 / Revised: 23 May 2014 / Accepted: 27 May 2014 / Published: 12 June 2014
(This article belongs to the Special Issue Oligosaccharides and Glyco-Conjugates)
Glycosaminoglycans (GAGs) due to their hydrophilic character and high anionic charge densities play important roles in various (patho)physiological processes. The identification and quantification of GAGs in biological samples and tissues could be useful prognostic and diagnostic tools in pathological conditions. Despite the noteworthy progress in the development of sensitive and accurate methodologies for the determination of GAGs, there is a significant lack in methodologies regarding sample preparation and reliable fast analysis methods enabling the simultaneous analysis of several biological samples. In this report, developed protocols for the isolation of GAGs in biological samples were applied to analyze various sulfated chondroitin sulfate- and hyaluronan-derived disaccharides using fluorophore-assisted carbohydrate electrophoresis (FACE). Applications to biologic samples of clinical importance include blood serum, lens capsule tissue and urine. The sample preparation protocol followed by FACE analysis allows quantification with an optimal linearity over the concentration range 1.0–220.0 µg/mL, affording a limit of quantitation of 50 ng of disaccharides. Validation of FACE results was performed by capillary electrophoresis and high performance liquid chromatography techniques.
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Keywords:
glycosaminoglycans; chondroitin sulfate; hyaluronan; fluorophore-assisted carbohydrate electrophoresis; capillary electrophoresis; high performance liquid chromatography
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MDPI and ACS Style
Karousou, E.; Asimakopoulou, A.; Monti, L.; Zafeiropoulou, V.; Afratis, N.; Gartaganis, P.; Rossi, A.; Passi, A.; Karamanos, N.K. FACE Analysis as a Fast and Reliable Methodology to Monitor the Sulfation and Total Amount of Chondroitin Sulfate in Biological Samples of Clinical Importance. Molecules 2014, 19, 7959-7980.
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