High-throughput analysis of immunoglobulin G glycosylation

ABSTRACT

Introduction: Glycosylation of immunoglobulin G (IgG) is important for its effector functions and was shown to be related to age, sex and disease status of an individual. Adding glycomic information to genome-wide association studies (GWAS) and large clinical trials is enabling insight into the functional relevance of changes in glycosylation, as well as molecular mechanisms behind these changes. Large-scale studies require sensitive, robust and affordable high-throughput methodologies for glycosylation analysis, which are currently available in only a limited number of laboratories.

Areas covered: This review focuses on currently used high-throughput approaches for N-glycosylation analysis of IgG, as well as some recent advances in the areas of deglycosylation, trypsin digestion, labeling, purification, derivatization and automation of current workflows. Relevant literature was searched using the PubMed database.

Expert commentary: Development, optimization and validation of robust, affordable and simple high-throughput glycosylation analysis methods is essential for discovery and validation of diagnostic and prognostic glycan biomarkers. Although significant advances in glycosylation analysis have been made in recent years, currently used protocols will have to be further optimized to enable subsequent analysis of glycosylation on all levels with the limited initial sample and in the minimal amount of time, which is still a challenging task.

KEYWORDS:

• Capillary electrophoresis
• glycan biomarker
• high-throughput
• IgG
• mass spectrometry
• N-glycosylation
• ultra-performance liquid chromatography

Acknowledgments

The authors would like to thank Dr. Thomas Stephen Klarić for critical reading of the manuscript.

Financial & competing interests disclosure

G. Lauc is the founder and owner of Genos Ltd, a private research organization that specializes in high-throughput glycomic analysis and has several patents in this field. I. Trbojević-Akmačić is the employee of Genos Ltd.

This research was supported by the European Commission FP7 projects HighGlycan (contract No. 278535) PainOmics (contract No. 602736), HTP-GlycoMet (contract No. 324400), and IntegraLife (contract No. 315997).