Recent advances in ion mobility–mass spectrometry for improved structural characterization of glycans and glycoconjugates

Abstract

Glycans and glycoconjugates are involved in regulating a vast array of cellular and molecular processes. Despite the importance of glycans in biology and disease, characterization of glycans remains difficult due to their structural complexity and diversity. Mass spectrometry (MS)-based techniques have emerged as the premier analytical tools for characterizing glycans. However, traditional MS-based strategies struggle to distinguish the large number of coexisting isomeric glycans that are indistinguishable by mass alone. Because of this, ion mobility spectrometry coupled to MS (IM-MS) has received considerable attention as an analytical tool for improving glycan characterization due to the capability of IM to resolve isomeric glycans before MS measurements. In this review, we present recent improvements in IM-MS instrumentation and methods for the structural characterization of isomeric glycans. In addition, we highlight recent applications of IM-MS that illustrate the enormous potential of this technology in a variety of research areas, including glycomics, glycoproteomics, and glycobiology.

Glycosylation of biosimilars: Recent advances in analytical characterization and clinical implications

“…Due to their mobility differences, the arrival time distribution can be measured and used to calculate the collision cross section (CCS). The CCS is a physical property related to the shape of the ion, representing a 3D fingerprint that can be used during characterization [121]. In addition to providing structural information, IM-MS data can significantly reduce the complexity of mass spectra by group separation in complex (biological) samples.…”

Glycans and glycosaminoglycans in neurobiology: key regulators of neuronal cell function and fate

“…While the structural complexity of glycan structures is a daunting subject to investigate [7][8][9][10] powerful analytics have been developed to assist in these investigations. These new methodologies include ion-mobility mass spectrometry [11,12], application of synchtrotron radiation for glycan structural analysis [13], application of high throughput automated N-glycopeptide glycoproteomic identification systems and orbitrap mass spectrometry [14][15][16], integrated systems glycobiology methodology incorporating glycogenomics, glycoproteomics and glycomics [17], fully automated chipelectrospray mass spectrometric analysis for the determination of CS/DS fine structure [18]. GAG microarrays for the analysis of GAG-protein interactions [19][20][21] have also been applied to profiling the sulphation patterns of GAGs to determine growth factor interactive sequences [22,23] and have also identified CS-E tetrasaccharides motifs which act as TNF antagonists [24].…”

Recommendations for Reporting Ion Mobility Mass Spectrometry Measurements

“…In 2006, the availability of IM-MS instrumentation from a major MS manufacturer marked the onset of a growing use of IM for the separation, identification, and structural characterization of analytes across diverse fields of science. These include: ionic clusters, [4][5][6][7][8] catalysts, 9,10 supramolecular complexes, [11][12][13][14] small organic molecules including drugs, 15,16 lipids 17,18 and other metabolites, [19][20][21] glycans, [22][23][24] peptides, [25][26][27] proteins, [28][29][30] synthetic polymers, [31][32][33] biomolecules 34,35 and biomolecular complexes [36][37][38] (native IM-MS). However, as IM-MS instruments now commercially available from different manufacturers operate according to different principles of ion mobility separation, 39 this introduces complexity-and potentially confusion-amongst practitioners and readers.…”