Work

De novo sequencing of glycans by ion mobility-mass spectrometry using a self-expanding database

Year: 2025

Type: article

Abstract: It is essential to determine glycan structures in complex biological samples to understand their biology and exploit their diagnostics, therapeutics and nutraceuticals potential. An unresolved analytical challenge is the identification of isomeric glycan structures in complex biological samples. Ion mobility (IM) combined with MS enables separation of isomeric glycans and identification by comparing their intrinsic collision cross section (CCS) values with similar data of synthetic standards. To identify glycans without the need to synthesize all biologically occurring glycans, we describe here an IM-MS de novo sequencing method based on fragment identification and sequence assembly. CCS values of additional fragments from glycans in biological samples result in a self-expanding reference database, gradually facilitating the sequencing of glycans of increasing complexity and expanding the database from an initial 19 standards to 332 unique entries. The methodology is employed to determine structures of human milk oligosaccharides and N-glycans of biotherapeutics.

Source: Nature Communications

DOI: 10.1038/s41467-025-67069-w