Utility of Ion-Mobility Spectrometry for Deducing Branching of Multiply Charged Glycans and Glycopeptides in a High-Throughput Positive ion LC-FLR-IMS-MS Workflow

作者：Edward G. Pallister, Matthew S. Choo, Ian Walsh, jien nee tai, Shi Jie Tay, Yuansheng Yang, Say Kong Ng, Pauline Mary Rudd, Sabine Lahja Flitsch, Terry Nguyen‐Khuong · 发表于：Analytical Chemistry · 年份：2020 · DOI：10.1021/acs.analchem.0c01954 · 被引用次数：42 · 研究领域：Glycosylation and Glycoproteins Research、Mass Spectrometry Techniques and Applications、Carbohydrate Chemistry and Synthesis

High-throughput glycan analysis has become an important part of biopharmaceutical production and quality control. However, it is still a significant challenge in the field of glycomics to easily deduce isomeric glycan structures, especially in a high-throughput manner. Ion mobility spectrometry (IMS) is an excellent tool for differentiating isomeric glycan structures. However, demonstrations of the utility of IMS in high-throughput workflows such as liquid chromatography-fluorescence-mass spectrometry (LC-FLR-MS) workflows have been limited with only a small amount of collision cross section (CCS) data available. In particular, IMS data of glycan fragments obtained in positive ion mode are limited in comparison to those obtained in negative ion mode despite positive ion mode being widely used for glycomics. Here, we describe IMS TW CCSN 2 data obtained from a high-throughput LC-FLR-IMS-MS workflow in positive ion mode. We obtained IMS data from a selection of RapiFluor-MS (RFMS) labeled N -glycans and also glycopeptides. We describe how IMS is able to distinguish isomeric N -glycans and glycopeptides using both intact IMS and fragment-based IMS glycan sequencing experiments in positive ion mode, without significantly altering the high-throughput nature of the analysis. For the first time, we were able to successfully use IMS in positive ion mode to determine the branching of isomeric glycopeptides and RFMS labeled glycans. Further, we highlight that IMS glycan sequencing of f...