al-Hakim A, Linhardt R J
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Electrophoresis. 1990 Jan;11(1):23-8. doi: 10.1002/elps.1150110106.
Acidic oligosaccharides derived from glycosaminoglycan heparin were separated by polyacrylamide gradient gel electrophoresis (PAGE). The gel could be visualized using Alcian Blue dye to give a pattern of highly resolved, well defined bands. The particular banding pattern obtained was the result of a heparinase catalyzed depolymerization which afforded oligosaccharide products that differed in size by one disaccharide unit. The separated oligosaccharides could be recovered prior to staining by electroelution onto a positively charged nylon membrane by a semi-dry transfer procedure. Subsequent elution and quantitative recovery of individual oligosaccharides from the membrane was achieved. By using multiple membrane layers a second separation dimension was obtained, resulting in increased oligosaccharide purity proportional to transfer depth. Preparative gradient polyacrylamide gel electrophoresis followed by semi-dry electro-transfer and recovery represents a novel method for the preparation of homogeneous acidic oligosaccharides.
从糖胺聚糖肝素衍生而来的酸性寡糖通过聚丙烯酰胺梯度凝胶电泳（PAGE）进行分离。使用阿尔辛蓝染料可使凝胶可视化，从而得到高度分辨、界限清晰的条带图谱。所获得的特定条带图谱是肝素酶催化解聚的结果，该解聚过程产生了大小相差一个二糖单位的寡糖产物。分离出的寡糖在染色前可通过半干转移程序电洗脱到带正电荷的尼龙膜上进行回收。随后实现了从膜上对单个寡糖的洗脱和定量回收。通过使用多个膜层获得了第二个分离维度，从而使寡糖纯度随转移深度增加。制备性梯度聚丙烯酰胺凝胶电泳，随后进行半干电转移和回收，代表了一种制备均一酸性寡糖的新方法。
