Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, Kawasaki, Kanagawa, Japan.
Methods Mol Biol. 2021;2271:169-178. doi: 10.1007/978-1-0716-1241-5_12.
O-glycosylation is a major post-translational modification of proteins. Accurate and detailed analysis to reveal O-glycosylation patterns at each site (site-specific O-glycosylation analysis) is essential to deeply understand glycoprotein function. Recent reports also demonstrated that unintended O-glycosylation occurs on therapeutic fusion glycoproteins; therefore, it is increasingly important to perform detailed and exhaustive O-glycosylation analysis during the development of therapeutic glycoproteins. Here, we describe a method of in-depth site-specific O-glycosylation analysis by liquid chromatography-mass spectrometry using electron-transfer/higher-energy collisional dissociation (EThcD) and database analysis.
O-糖基化是蛋白质的一种主要翻译后修饰。准确详细的分析以揭示每个位点的 O-糖基化模式（位点特异性 O-糖基化分析）对于深入了解糖蛋白的功能至关重要。最近的报告还表明，治疗性融合糖蛋白上会发生意外的 O-糖基化；因此，在治疗性糖蛋白的开发过程中，进行详细和详尽的 O-糖基化分析变得越来越重要。在这里，我们描述了一种使用液相色谱-质谱联用技术通过电子转移/更高能量碰撞解离（EThcD）和数据库分析进行深入的位点特异性 O-糖基化分析的方法。
