用于测定糖蛋白碳水化合物部分的液相色谱法。应用于α1-酸性糖蛋白和组织型纤溶酶原激活剂。
Liquid chromatographic method for the determination of the carbohydrate moiety of glycoproteins. Application to alpha 1-acid glycoprotein and tissue plasminogen activator.
作者信息
Taverna M, Baillet A, Werner R, Baylocq-Ferrier D
机构信息
Centre d'Etudes Pharmaceutiques, Laboratoire de Chimie Analytique, Chatenay Malabry, France.
出版信息
J Chromatogr. 1991 Sep 27;558(1):105-14. doi: 10.1016/0021-9673(91)80115-w.
A rapid procedure is described for the qualitative and quantitative analysis of the carbohydrate composition of glycoproteins by liquid chromatography with light-scattering detection. The analysis was carried out in three steps. First, the glycoprotein samples were purified by a two-step purification on a Sephadex G-25 column with a 90% yield. Second, the selectivity of the separation and the sensitivity of detection of monosaccharides, as methyl glycosides obtained by direct methanolysis of glycoproteins, were improved by modified simplex optimization of the methanolysis parameters (temperature, methanolic hydrochloric acid strength and reaction time) determined at 66 degrees C, 1.2 M and 8.1 h for alpha 1-acid glycoprotein (alpha-AGP) and 73 degrees C, 1.5 M and 12.5 h for tissue plasminogen activator (tPA). Finally, the method was applied to the determination of the carbohydrate moiety of the two N-glycosylated glycoproteins alpha-AGP and tPA.
本文描述了一种通过液相色谱-光散射检测对糖蛋白碳水化合物组成进行定性和定量分析的快速方法。该分析分三步进行。首先，糖蛋白样品在Sephadex G-25柱上通过两步纯化，产率为90%。其次，通过对糖蛋白直接甲醇解得到的甲基糖苷形式的单糖进行分离选择性和检测灵敏度的优化，对甲醇解参数（温度、甲醇盐酸浓度和反应时间）进行改进的单纯形优化，对于α1-酸性糖蛋白（α-AGP），在66℃、1.2M和8.1小时条件下确定参数；对于组织纤溶酶原激活剂（tPA），在73℃、1.5M和12.5小时条件下确定参数。最后，该方法应用于两种N-糖基化糖蛋白α-AGP和tPA的碳水化合物部分的测定。
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