通过 LC/MS 进行重组蛋白的位点特异性 N-糖基化分析。

Site-Specific N-glycosylation Analysis of Recombinant Proteins by LC/MS.

机构信息

SGS M-Scan SA, Plan-les-Ouates, Switzerland.

出版信息

Methods Mol Biol. 2021;2271:133-154. doi: 10.1007/978-1-0716-1241-5_10.

Abstract

The glycosylation process is extremely heterogeneous, dynamic, and complex compared with any other post-translational modification of protein. In the context of recombinant glycoproteins, glycosylation is a critical attribute as glycans could dramatically alter protein functions and properties including activity, half-life, in vivo localization, stability, and, last but not least, immunogenicity. Liquid chromatography combined to mass spectrometry constitutes the most powerful analytical approach to achieve the comprehensive glycan profile description or comparison of glycoproteins. This chapter details a versatile yet straightforward LC-MS approach for sample preparation, analysis, and data interpretation, enabling the evaluation of site-specific N-glycosylation of recombinant glycoproteins.

摘要

与蛋白质的任何其他翻译后修饰相比，糖基化过程非常复杂、动态且具有异质性。在重组糖蛋白的情况下，糖基化是一个关键属性，因为聚糖可以极大地改变蛋白质的功能和特性，包括活性、半衰期、体内定位、稳定性，最后但并非最不重要的是免疫原性。液相色谱与质谱联用是实现糖蛋白全面聚糖谱描述或比较的最强大的分析方法。本章详细介绍了一种通用且简单的 LC-MS 方法，用于样品制备、分析和数据解释，从而能够评估重组糖蛋白的位点特异性 N-糖基化。