Recent advances in the capillary electrophoresis of recombinant glycoproteins.
Recent advances in the capillary electrophoresis of recombinant glycoproteins.
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DOI：
10.1016/s0003-2670(98)00495-4
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发表时间：
1999-03
影响因子：
6.2
通讯作者：
A. Pantazaki;M. Taverna;C. Vidal-madjar
中科院分区：
文献类型：
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作者：
A. Pantazaki;M. Taverna;C. Vidal-madjar
需要高效的方法来分析用于临床用途的重组蛋白。这些蛋白质通常由哺乳动物表达系统产生，是高度糖基化的，并且由糖基化变体（糖型）群体组成。本文综述了毛细管电泳（CE）用于完整重组糖蛋白分析和监测其生物生产的各种微尺度技术。毛细管区带电泳（CZE）具有操作简便、快速、自动化程度高等优点，已被广泛应用于完整糖蛋白糖型群的分析。毛细管等电聚焦（CIEF）是一种基于电荷的糖型分离的有效方法。胶束电动毛细管色谱法（MEKC）代表了CZE的替代方法，用于重组糖蛋白的纯度控制，而十二烷基硫酸钠-毛细管凝胶电泳（SDS-CGE）与可更换的凝胶基质给出了糖型分子量的估计。CIEF和SDS-CGE的结果与相应的平板凝胶技术的结果相当。CZE与质谱（MS）耦合的最新进展不仅为精确的分子量测定提供了新的视角，而且还为更好地理解糖型的CE分离机制提供了新的视角。
Highly efficient methods are required to analyze recombinant proteins for clinical use. These proteins generally produced from mammalian expression systems are highly glycosylated and consist of a population of glycosylated variants (glycoforms). This review presents the different microscale techniques of capillary electrophoresis (CE) for analyzing the intact recombinant glycoproteins and for monitoring their bioproduction. Because of several advantages such as simplicity, speed and automation, capillary zone electrophoresis (CZE) has been generally employed for the routine analysis of the glycoform populations of intact glycoproteins. Capillary isoelectric focusing (CIEF) is a powerful method for a charge-based separation of the glycoforms. Micellar electrokinetic capillary chromatography (MEKC) represents an alternative method to CZE for the purity control of recombinant glycoproteins, while the sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) with replaceable gel matrices gives an estimation of the glycoform molecular masses. The results from CIEF and SDS-CGE are comparable to those from the corresponding slab gel techniques. The recent advances in the coupling of CZE with mass-spectrometry (MS) offers new perspectives not only for precise molecular mass determinations, but also to better understand the mechanisms involved in the CE separation of glycoforms.