Advances in analysis of glycosaminoglycans: its application for the assessment of physiological and pathological states of connective tissues

Vynios, Demitrios H.; Karamanos, Nikos K.; Tsiganos, Constantine P.

doi:10.1016/s1570-0232(02)00498-1

Cited by 51 publications

(56 citation statements)

References 114 publications

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“…Treatment of CS isomers with chondro/dermato lyases is essential to determine disaccharides composition. The obtained D-disaccharides have been previously measured by either HPLC or CE by UVdetection at 232 nm due to the absorbance of the unsaturated disaccharide released by chondroitinases [21][22][23][24][25]. However, since plasma GAGs are found in trace amounts, highly sensitive methods are required for their analysis and structural characterization.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2007

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The report describes a rapid and simple CE method using LIF detection for the analysis of unsaturated disaccharides obtained from enzymatic depolymerization of plasma chondroitin sulfate (CS) isomers. The disaccharide reducing groups were labeled with 2-aminoacridone (AMAC). The fluorotagged products can be separated by reversed-polarity CE using a sodium acetate buffer, pH 3.8, in the presence of 0.05% methylcellulose. The choice of the appropriate electrophoretic conditions was performed after a deep analysis of the most important parameters affecting analyte separation. In particular, the effect of both run buffer concentration and pH on resolution, efficiency, migration times, and peak area was evaluated. The selected electrophoretic conditions allowed us to separate the CS isomers-derived Delta-disaccharides in less than 12 min, also resolving the nonsulfated disaccharides released from CS isomers from those released from hyaluronan (HA). Moreover, these conditions gave a good reproducibility of both the migration times (CV%, 0.25) and the peak areas (CV%, 1.4). Intra- and interassay CV were 5.37 and 7.23%, respectively, and analytical recovery was about 86%. The applicability of the above method to the quantitative and structural disaccharide analyses of plasma CS isomers was investigated. Data obtained from 44 healthy human subjects were compared with those obtained by a fluorophore-assisted carbohydrate electrophoresis (FACE) reference assay, by using the Passing and Bablok regression and Bland-Altman tests. The developed method could represent a good tool for an ultrasensitive analysis of CS isomers in biological samples from different sources, particularly when samples are available in very low amounts.

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Section: Introductionmentioning

confidence: 99%

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

et al. 2007

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“…GAGs are highly anionic polysaccharides present in the extracellular matrix (ECM) and the cells. The biological importance of their fine structures in normal and disease state has been well demonstrated (1). They seem to play several roles in tumor formation because of their capacity to bind and modulate the action of a variety of proteins including growth factors, cytokines, and proteases.…”

Section: Introductionmentioning

confidence: 99%

Alterations of glycosaminoglycan disaccharide content and composition in colorectal cancer: Structural and expressional studies

Kalathas

Theocharis²,

Bounias³

et al. 2009

Oncol Rep

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Abstract.The glycosaminoglycans are implicated in many processes important in the growth and progression of malignant tumors. In the present study glycosaminoglycans were purified from healthy, macroscopically normal and cancerous specimens of different anatomic sites and different stages of cancer and analyzed by FACE after chondroitinases and sulfatases digestion. The cancerous samples contained increased levels of 6-sulfated unsaturated disaccharides compared to macroscopically normal and healthy samples, the increase being stage-related. The differences in sulfation were found to be related to the anatomic site and the stage of cancer. RT-PCR analysis of 4-sulfotransferase mRNA revealed its presence in decreasing amounts as the stage of the cancer increased. Furthermore, the percent content of hyaluronan disaccharides was elevated in macroscopically normal samples compared to the cancerous, and in addition, it was much more elevated than that of healthy samples. Haluronan levels increase with stage in cancerous tissues. Therefore, it could be concluded that the glycosaminoglycans in colorectal cancer are biosynthetically directed to contribute in different ways depending on the cancer stage and anatomical site.

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“…Together with heparin, heparin sulfate, dermatan sulfate, keratan sulfate, and chondroitin sulfate (CS), it forms the group of connective tissue polysaccharides known as glycosaminoglycans (GAGs). HA has a high molecular mass (up to 8610 6 Da corresponding to about 10 000 disaccharide units), is not sulfated, and is not covalently bound to proteins [1][2][3][4][5][6]. HA is mainly produced by fibroblasts and other specialized connective tissue cells and participates in various cell-to-cell interactions.…”

Section: Introductionmentioning

confidence: 99%

“…Monitoring serum HA is also used in other areas, such as in septic conditions, tumor growth (such as Wilm's tumor and malignant mesothelioma), and for the diagnosis of rare hereditary diseases (such as Werner's syndrome and cutaneous hyaluronanosis) [4]. Many different colorimetric, chromatographic, and electrophoretic methods have been developed for the analysis of HA and other GAGs, their metabolic products, and the degraded molecules thereof [1,2]. Serum HA is currently determined with assays that are based upon the specific affinity of certain proteins to HA and that feature a low lg/L sensitivity [17], most notably a radiometric method (HA-test, Pharmacia, Uppsala, Sweden) and a sandwich enzyme-binding assay (Chugai Pharmaceutical, Tokyo, Japan; HA-test, Corgenix, Westminster, CO, USA).…”

Section: Introductionmentioning

confidence: 99%

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

Yang

Breadmore

Thormann³

2005

J of Separation Science

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Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking CE conditions for monitoring the unsaturated disaccharides of hyaluronic acid (di-HA) and chondroitin sulfate (di-CS) using an alkaline tetraborate buffer, electrokinetic sample injection, and UV absorption detection at 232 nm are reported. Separations were performed in an uncoated fused-silica capillary having reversed polarity and reversed electroosmosis generated with the addition of CTAB to the buffer. The influence of various separation parameters, including the concentration of CTAB, buffer pH, concentration of tetraborate, and applied voltage, on the resolution of the two disaccharides was investigated. Baseline separation was obtained with 25 mM tetraborate at pH 10.0 and having 0.05 mM CTAB. Chloride and phosphate in the sample are beneficial for the stacking of the disaccharides, with di-HA forming a much sharper peak than di-CS. Using samples prepared in 25 mM Tris-HCl (pH 7.5) and electrokinetic injection at the cathode at -10 kV for 40 s, linear relationships between the corrected peak area and the concentration of the disaccharides have been found in the ranges of 1.0-400.0 and 0.1-1.0 lg/mL (0.2-1.0 lg/mL for di-CS), with correlation coefficients being A0.9933 in all cases. The RSDs of detection times and corrected peak areas were between 1.13-1.24 and 1.57-2.13%, respectively. Applied to human serum samples that were prepared by ethanol precipitation and depolymerization of the two polysaccharides with chondroitinase ABC reveals comigration of endogenous compounds with di-HA and a sample-dependent detection time. The di-HA content in the serum sample can be estimated via subtraction of the blank peak that is obtained without enzymatic hydrolysis.

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Advances in analysis of glycosaminoglycans: its application for the assessment of physiological and pathological states of connective tissues

Cited by 51 publications

References 114 publications

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

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

Alterations of glycosaminoglycan disaccharide content and composition in colorectal cancer: Structural and expressional studies

Analysis of the disaccharides derived from hyaluronic acid and chondroitin sulfate by capillary electrophoresis with sample stacking

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