Comparative performance of four methods for high-throughput glycosylation analysis of immunoglobulin G in genetic and epidemiological research

The biological and clinical relevance of glycosylation is becoming increasingly recognised, 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 (CGE), and two mass spectrometric (MS) methods for quantitative profiling of N-glycosylation of IgG in the same dataset of 1201 individuals. To evaluate 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 CGE, 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.