Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore; NUS Environmental Research Institute, 5A Engineering Drive 1, T-Lab Building, Singapore 117411, Singapore.
Department of Chemistry, National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore.
Anal Chim Acta. 2017 Feb 8;953:79-86. doi: 10.1016/j.aca.2016.11.043. Epub 2016 Nov 25.
The deep involvement of glycans or carbohydrate moieties in biological processes makes glycan patterns an important direction for the clinical and medicine researches. A multiplexing CE mapping method for glycan analysis was developed in this study. By applying different CE separation mechanisms, the potential of combined parallel applications of capillary zone electrophoresis (CZE), micellar electrokinetic chromatography (MEKC) and capillary gel electrophoresis (CGE) for rapid and accurate identification of glycan was investigated. The combination of CZE and MEKC demonstrated enhancing chromatography separation capacity without the compromises of sample pre-treatment and glycan concentration. The separation mechanisms for multiplexing platform were selected based on the orthogonalities of the separation of glycan standards. MEKC method exhibited promising ability for the analysis of small GU value glycans and thus complementing the unavailability of CZE. The method established required only small amount of samples, simple instrument and single fluorescent labelling for sensitive detection. This integrated method can be used to search important glycan patterns appearing in biopharmaceutical products and other glycoproteins with clinical importance.
聚糖或碳水化合物部分在生物过程中的深度参与使糖型成为临床和医学研究的一个重要方向。本研究开发了一种用于聚糖分析的多重 CE 图谱方法。通过应用不同的 CE 分离机制，研究了毛细管区带电泳（CZE）、胶束电动色谱（MEKC）和毛细管凝胶电泳（CGE）的组合平行应用在快速准确鉴定聚糖方面的潜力。CZE 和 MEKC 的组合证明了在不影响样品预处理和聚糖浓度的情况下增强了色谱分离能力。基于聚糖标准的分离正交性选择了多重平台的分离机制。MEKC 方法在分析小 GU 值聚糖方面表现出良好的能力，从而弥补了 CZE 的不足。所建立的方法仅需要少量样品、简单的仪器和单一的荧光标记即可进行灵敏检测。这种集成方法可用于搜索生物制药产品和其他具有临床意义的糖蛋白中出现的重要糖型模式。
