Irimura T, Nakajima M, Di Ferrante N, Nicolson G L
Anal Biochem. 1983 Apr 15;130(2):461-8. doi: 10.1016/0003-2697(83)90616-4.
A high-speed gel-permeation chromatographic system for analyzing glycosaminoglycans which uses two 0.7 X 75-cm stainless-steel columns containing Fractogel (Toyopearl) TSK HW-55(S), was developed. Glycosaminoglycans were applied and eluted with a 0.2 M sodium chloride solution and monitored by ultraviolet absorption at 210 nm or radioactivity. The best resolution of glycans was obtained at 55 degrees C at a flow rate of 1.0 ml/min. Acidic and neutral glycans in the molecular weight (Mr) range 600-60,000 eluted within 45 min. A linear relationship was found between retention time and molecular weight using standard glycosaminoglycans, chitin oligosaccharides, and a porcine thyroglobulin glycoprotide. This system was used to analyze the heparan sulfate synthesized by PYS-2 embryonic carcinoma cells and the degradation products produced by incubating it with extracted glycosidases from metastatic B16 melanoma cells. The results indicated that B16 melanoma cells contain at least two different heparan sulfate degradative activities, one of which appears to be an endoglycosidase.
开发了一种用于分析糖胺聚糖的高速凝胶渗透色谱系统，该系统使用两根0.7×75厘米的不锈钢柱，柱内填充Fractogel（Toyopearl）TSK HW - 55(S)。糖胺聚糖用0.2 M氯化钠溶液进样和洗脱，并通过210 nm处的紫外吸收或放射性进行监测。在55℃、流速为1.0 ml/min时，聚糖获得了最佳分离效果。分子量（Mr）在600 - 60,000范围内的酸性和中性聚糖在45分钟内洗脱完毕。使用标准糖胺聚糖、几丁质寡糖和猪甲状腺球蛋白糖蛋白发现保留时间与分子量之间存在线性关系。该系统用于分析PYS - 2胚胎癌细胞合成的硫酸乙酰肝素以及将其与转移性B16黑色素瘤细胞提取的糖苷酶一起孵育产生的降解产物。结果表明，B16黑色素瘤细胞至少含有两种不同的硫酸乙酰肝素降解活性，其中一种似乎是内切糖苷酶。
