Comparative Performance of Four Methods for High-throughput Glycosylation Analysis of Immunoglobulin G in Genetic and Epidemiological Research

Abstract

The biological and clinical relevance of glycosylation is becoming increasingly recognized, leading to a growing interest in large-scale clinical and population-based studies. In the past few years, several methods for high-throughput analysis of glycans have been developed, but thorough validation and standardization of these methods is required before significant resources are invested in large-scale studies. In this study, we compared liquid chromatography, capillary gel electrophoresis, and two MS methods for quantitative profiling of N-glycosylation of IgG in the same data set of 1201 individuals. To evaluate the accuracy of the four methods we then performed analysis of association with genetic polymorphisms and age. Chromatographic methods with either fluorescent or MS-detection yielded slightly stronger associations than MS-only and multiplexed capillary gel electrophoresis, but at the expense of lower levels of throughput. Advantages and disadvantages of each method were identified, which should inform the selection of the most appropriate method in future studies.

Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes

“…For each participant, N-glycans were released from plasma proteins, fluorescently labelled and analysed by HILIC-UPLC, a method demonstrated to be the best approach for reliable and reproducible quantitative glycan analysis [14]. By this method, plasma glycome is separated into 46 chromatographic peaks, each containing a group of similar glycan structures.…”

Human Plasma N-glycosylation as Analyzed by Matrix-Assisted Laser Desorption/Ionization-Fourier Transform Ion Cyclotron Resonance-MS Associates with Markers of Inflammation and Metabolic Health

“…Glycan compositions as obtained by MS provide orthogonal information to chromatographic peaks. UHPLC, for example, can separate diantennary N-glycan isomers with ␣1,3-versus ␣1,6-arm galactosylation and can likewise distinguish an antennary from a bisecting GlcNAc, whereas MS can provide more precise groups of di-, tri-, and tetraantennary compositions, number of sialic acids, and clear separation of highmannose type glycans (29,71). As such, the construction of derived glycosylation traits making use of these features, while still biased on a compositional level, is simple to perform and provides additional insight into the complexity of glycan changes.…”

Glycosylation Profile of IgG in Moderate Kidney Dysfunction

“…38 Approximately 50% of IgG glycans are not sialylated and are proinflammatory. 39 However, the terminal a2,6-sialylation of IgG glycans decreases the ability of IgG to bind Fcg receptors (FcgRs), which increases expression of inhibitory FcgRIIB and is anti-inflammatory. 40 Contrary to changes in galactosylation, the significant changes in sialylation have not been associated with other diseases.…”