Waters Corporation, Milford, MA, USA.
GlaxoSmithKline, Stevenage, UK.
Methods Mol Biol. 2021;2271:205-219. doi: 10.1007/978-1-0716-1241-5_15.
Analysis of N-glycans are commonly conducted via enzymatic release, labeling, and liquid chromatography (LC) separation and fluorescent detection. Mass spectrometry (MS) has been increasingly used as an orthogonal detection method to provide additional structural information and increase the confidence of N-glycan analysis. In this chapter, we describe a method to perform routine analysis of N-glycans including the sample preparation with a signal-enhancement label, LC-MS data generation, and data analysis. Using this method, up to 24 N-glycan samples can be prepared at one time and analyzed by LC-MS. With the addition of automation platform, up to 96 N-glycan samples can be prepared and analyzed in a high-throughput manner.
N-糖链分析通常通过酶解释放、标记、液相色谱 (LC) 分离和荧光检测来进行。质谱 (MS) 已越来越多地被用作正交检测方法，以提供额外的结构信息并提高 N-糖链分析的可信度。在本章中，我们描述了一种常规分析 N-糖链的方法，包括带有信号增强标签的样品制备、LC-MS 数据生成和数据分析。使用这种方法，一次最多可以制备 24 个 N-糖链样品并进行 LC-MS 分析。通过添加自动化平台，一次最多可以制备和分析 96 个 N-糖链样品，实现高通量分析。
