Routine Analysis of N-Glycans Using Liquid Chromatography Coupled to Routine Mass Detection

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.

Analytical characterization; Automation; Cell line development; Critical quality attribute; High-throughput analysis; LC-MS; N-glycan analysis; Process development; Routine mass detection.