Karamanos N K, Vanky P, Tzanakakis G N, Hjerpe A
Department of Immunology, Microbiology, Pathology and Infectious Diseases, Huddinge University Hospital F-42, Sweden.
Electrophoresis. 1996 Feb;17(2):391-5. doi: 10.1002/elps.1150170217.
A rapid, sensitive and accurate high-performance capillary electrophoresis method is described for the determination of the sulfation pattern of heparin and heparan sulfate disaccharides. The analysis, performed after enzymic degradation of the polysaccharides with heparinase and heparinases II and III in combination, yields highly UV-absorbing delta-disaccharides. The separation is performed with reversed polarity using 15 mM phosphate buffer, pH 3.50. This method is superior to others since all known 12 disaccharides carrying N-acetylated, N-sulfated or unsubstituent glucosamine can be separated in a single run of 15 min. At the highest sensitivity the analysis consumes only a few femtograms of glycosaminoglycan and allows a determination of delta-disaccharides at the attomole level.
本文描述了一种快速、灵敏且准确的高效毛细管电泳方法，用于测定肝素和硫酸乙酰肝素二糖的硫酸化模式。在用肝素酶、肝素酶II和III联合对多糖进行酶解后进行分析，可产生高紫外吸收的δ-二糖。使用15 mM pH 3.50的磷酸盐缓冲液以反相模式进行分离。该方法优于其他方法，因为所有已知的携带N-乙酰化、N-硫酸化或未取代葡糖胺的12种二糖均可在15分钟的单次运行中分离出来。在最高灵敏度下，该分析仅消耗几飞克的糖胺聚糖，并能在阿托摩尔水平测定δ-二糖。
