Jackson Thomas R, Springall Jeremy S, Rogalle Damien, Masumoto Naoko, Ching Li Hung, D'Hooge François, Perera Semali P, Jenkins A Toby A, James Tony D, Fossey John S, van den Elsen Jean M H
Department of Biology and Biochemistry, University of Bath, Bath, UK.
Electrophoresis. 2008 Nov;29(20):4185-91. doi: 10.1002/elps.200800178.
The incorporation of specialised carbohydrate affinity ligand methacrylamido phenylboronic acid in polyacrylamide gels for fluorophore-assisted carbohydrate electrophoresis greatly improved the effective separation of saccharides that show similar mobilities in standard electrophoresis. Polyacrylamide gel electrophoresis using methacrylamido phenylboronic acid in low loading (typically 0.5-1% dry weight) was unequivocally shown to alter retention of labelled saccharides depending on their boronate affinity. While conventional fluorophore-assisted carbohydrate electrophoresis of 2-aminoacridone labelled glucose oligomers showed an inverted parabolic migration, an undesired trait of small oligosaccharides labelled with this neutral fluorophore, boron affinity saccharide electrophoresis separation of these carbohydrates completely restored their predicted running order, based on their charge/mass ratio, and resulted in improved separation of the analyte saccharides. These results exemplify boron affinity saccharide electrophoresis as an important new technique for analysing carbohydrates and sugar-containing molecules.
在用于荧光团辅助碳水化合物电泳的聚丙烯酰胺凝胶中加入特殊的碳水化合物亲和配体甲基丙烯酰胺基苯硼酸，极大地改善了在标准电泳中迁移率相似的糖类的有效分离。使用低负载量（通常为干重的0.5 - 1%）的甲基丙烯酰胺基苯硼酸进行聚丙烯酰胺凝胶电泳，明确显示出根据标记糖类的硼酸亲和力改变其保留情况。虽然传统的2-氨基吖啶酮标记的葡萄糖寡聚物的荧光团辅助碳水化合物电泳显示出倒抛物线形迁移，这是用这种中性荧光团标记的小寡糖的一个不良特性，但基于电荷/质量比，这些碳水化合物的硼亲和糖类电泳分离完全恢复了它们预测的运行顺序，并改善了分析物糖类的分离。这些结果例证了硼亲和糖类电泳是分析碳水化合物和含糖分子的一项重要新技术。
