生物样品中荧光团标记的透明质酸和硫酸软骨素二糖的分析。

Analysis of fluorophore-labelled hyaluronan and chondroitin sulfate disaccharides in biological samples.

作者信息

Karousou Evgenia G, Porta Giovanni, De Luca Giancarlo, Passi Alberto

机构信息

Department of Experimental and Clinical Biomedical Sciences, University of Insubria, Varese, Italy.

出版信息

J Pharm Biomed Anal. 2004 Mar 1;34(4):791-5. doi: 10.1016/S0731-7085(03)00568-5.

DOI:10.1016/S0731-7085(03)00568-5

PMID:15019056

Abstract

In this report, we describe a polyacrylamide gel electrophoresis for the analysis of fluorophore-labelled hyaluronan (HA) and chondroitin sulfate (CS) Delta-disaccharides. The method utilizes derivatization of reducing ends of hyaluronan and the variously sulfated chondroitin sulfate Delta-disaccharides with 2-aminoacridone (AMAC), followed by electrophoresis in Tris-HCl buffer (pH 8.8), and polyacrylamide gel (T 25%/C 3.75%). The method was applied to the analysis of GAGs secreted into the culture medium of human aortic smooth muscle cells and the obtained results were compared with those analysed by fluorophore assisted carbohydrate electrophoresis (FACE). The described method is a useful and sensitive tool for the rapid monitoring of GAGs in high number of biologic samples.

摘要

在本报告中，我们描述了一种用于分析荧光团标记的透明质酸（HA）和硫酸软骨素（CS）Δ-二糖的聚丙烯酰胺凝胶电泳方法。该方法利用2-氨基吖啶酮（AMAC）对透明质酸的还原端以及各种硫酸化的硫酸软骨素Δ-二糖进行衍生化，随后在Tris-HCl缓冲液（pH 8.8）和聚丙烯酰胺凝胶（T 25%/C 3.75%）中进行电泳。该方法应用于分析分泌到人主动脉平滑肌细胞培养基中的糖胺聚糖，并将所得结果与通过荧光团辅助碳水化合物电泳（FACE）分析的结果进行比较。所描述的方法是一种用于快速监测大量生物样品中糖胺聚糖的有用且灵敏的工具。