A survey of methodological challenges for glycosaminoglycan/proteoglycan analysis and structural characterization by capillary electrophoresis

Karamanos, Nikos K.; Hjerpe, Anders

doi:10.1002/elps.1150191504

Cited by 48 publications

(29 citation statements)

References 57 publications

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“…Application of CE to carbohydrate analysis has offered considerable information due to the high sensitivity of the technique and the great resolving power. The advances in the field have been extensively recorded in a number of recent reviews (ElRassi and Merchef, 1996;Honda, 1996;Kakehi and Honda 1996;Linhardt and Pervin, 1996;Novotny, 1996;Paulus and Klockow, 1996;Karamanos and Hjerpe, 1998;Lamari and Karamanos, 1999).…”

Section: Application Of Ce In the Area Of Glycoconjugates Introductormentioning

confidence: 99%

State-of-the-art of capillary electrophoresis with application to the area of glycoconjugates

Karamanos

Lamari

1999

Biomed. Chromatogr.

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Glycoconjugates constitute a major group of biomolecules, which participate in several biological functions and processes. Their carbohydrate components play key roles in determining the properties of glycoconjugates and, therefore, analysis and structural characterization of carbohydrates are essential. Capillary electrophoresis, due to its high resolving power and sensitivity, has been successfully used for the analysis of carbohydrates. In this review the principles of high‐performance capillary electrophoresis; mechanisms employed for glycoconjugate analysis as well as the various detection techniques used are summarized. Copyright © 1999 John Wiley & Sons, Ltd.

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Section: Application Of Ce In the Area Of Glycoconjugates Introductormentioning

confidence: 99%

State-of-the-art of capillary electrophoresis with application to the area of glycoconjugates

Karamanos

Lamari

1999

Biomed. Chromatogr.

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“…Capillary electrophoresis (CE) with its high resolving power and sensitivity is a promising technique for the analysis of glycan/PG (for reviews see Grimshaw, 1997;Karamanos and Hjerpe, 1998b). Various CE methods and strategies have been used for determining GAG/PG structures.…”

Section: Strategies For Analysis Of Glycans/ Proteoglycans Ce Analysimentioning

confidence: 99%

Strategies for analysis and structure characterization of glycans/proteoglycans by capillary electrophoresis. Their diagnostic and biopharmaceutical importance

Karamanos

Hjerpe

1999

Biomed. Chromatogr.

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Proteoglycans are key biological macromolecules that, via their glycan constituents, participate and regulate several cellular events and physiopathological processes. Refined structures of their highly anionic glycan chains, involving sulphation pattern and uronic acid distribution through the polymeric chain, determine the interactions of proteoglycans with matrix effector molecules and are responsible for numerous effects. Analysis and structural characterization of glycans are, therefore, essential in understanding the biological functions of proteoglycans. Capillary electrophoresis with its high resolving power and sensitivity may successfully be used for the fine chemical characterization of components present in low amounts and to overcome limitations due to low amount/volume of biologic samples available. This technique is also friendly to the user and to the environment, since a very small amount of solvents (a few µL) is required. In this review the strategies used to analyse and characterize the structure of glycan chains of proteoglycans are summarized. They involve capillary electrophoretic analysis of depolymerized acidic glycan chains using specific enzymes and analysis of intact chains. The importance of this type of analysis in biologic samples and tissues and its possible diagnostic and biopharmaceutical use are also discussed. Copyright © 1999 John Wiley & Sons, Ltd.

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“…Thus, HPCE has extended its applicability to various kinds of carbohydrates, including monosaccharides, oligosaccharides, glycosaminoglycans (GAGs), and glycans of glycoproteins. [5][6][7][8][9][10] These methods are superior in small sample consumption and easy operations to other methods including HPLC analyses.…”

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Separation of 2-Aminobenzoic Acid-Derivatized Glycosaminoglycans and Asparagine-linked Glycans by Capillary Electrophoresis

Sato

Ōkubo

et al. 2005

ANAL. SCI.

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Glycosylation is known to be the most common and diverse post-translational modification of proteins. It may affect their biological activity, solubility, stability, and immunogenicity. Therefore, determining the sugar structures has become increasingly important to understand and elucidate the functions of these molecules.To analyze carbohydrates, a high separation efficiency is required, because oligosaccharides have branched structures and comprise a variety of isomers and homologues. Chromatographic methods have recently been commonly used to identify the oligosaccharide structures in a glycoprotein. They are powerful methods for the structural characterization of N-linked oligosaccharides, and consume only picomoles of samples. For example, a two-dimensional sugar mapping technique for pyridylaminated oligosaccharides 1-3 or a highperformance anion-exchange chromatography with a pulsed amperometric detector 4 are well known as high-resolution and sensitive methods.High-performance capillary electrophoresis (HPCE), which is a relatively new separation technique, is known as a highresolution, simple and rapid method. Usually, it does not require any pretreatment steps, and permits analyses by a variety of separation modes simply by changing the selection of a running electrolyte solution. Thus, HPCE has extended its applicability to various kinds of carbohydrates, including monosaccharides, oligosaccharides, glycosaminoglycans (GAGs), and glycans of glycoproteins.5-10 These methods are superior in small sample consumption and easy operations to other methods including HPLC analyses.Because most of carbohydrates do not have strong absorbance in UV and visible regions, labeling procedures are necessary for HPCE analyses. 2-Aminobenzoic acid (2-AA) is a useful derivatizing reagent for carbohydrate analyses. [11][12][13][14][15][16][17][18] The 2-AA labeling procedure is rapid, selective, and easy to perform. 14 Moreover, 2-AA derivatives are able to become charged positively, negatively, or neutral, depending on the pH of a running buffer, since it has both a carboxyl group and an amino group. 15 This charge flexibility is suitable for multidimensional HPCE analysis. We have reported on a highly efficient derivatizing method and the determination of 2-AA-derivatized monosaccharides by HPCE with UV detection. 15,17,18 This method has advantages of simplicity, rapidness, high resolution, and high sensitivity. In the present paper, we report that the HPCE separations of 2-AA-derivatized GAGs and N-linked glycans of glycoproteins have also been achieved. Experimental ReagentsAll of the reagents were of the highest grade available and used without any further purification. Sodium cyanoborohydride was from Aldrich. Various sulfated heparinderived-∆-disaccharides and heparan sulfate-derived-∆-disaccharides, chondroitin sulfate-∆-disaccharides, hyaluronic acid, keratan sulfate from bovine cornea, glucose-oligomer, Nglycan, chondroitinase ABC, and keratenase II (Bacillus sp.) were all purchased from Seikagaku Kogyo Corp....

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A survey of methodological challenges for glycosaminoglycan/proteoglycan analysis and structural characterization by capillary electrophoresis

Cited by 48 publications

References 57 publications

State-of-the-art of capillary electrophoresis with application to the area of glycoconjugates

State-of-the-art of capillary electrophoresis with application to the area of glycoconjugates

Strategies for analysis and structure characterization of glycans/proteoglycans by capillary electrophoresis. Their diagnostic and biopharmaceutical importance

Separation of 2-Aminobenzoic Acid-Derivatized Glycosaminoglycans and Asparagine-linked Glycans by Capillary Electrophoresis

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