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MIgGGly (mouse IgG glycosylation analysis) - a high-throughput method for studying Fc-linked IgG N-glycosylation in mice with nanoUPLC-ESI-MS

Zaytseva, Olga; Jansen, Bas; Hanić, Maja; Mrčela, Mia; Razdorov, Genadij; Stojković, Ranko; Erhardt, Julija; Brizić, Ilija; Jonjić, Stipan; Pezer, Marija; Lauc, Gordan (2018) MIgGGly (mouse IgG glycosylation analysis) - a high-throughput method for studying Fc-linked IgG N-glycosylation in mice with nanoUPLC-ESI-MS. Scientific Reports, 8 . ISSN 2045-2322

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Abstract

Immunoglobulin G (IgG) N-glycosylation is crucial for its effector functions. It is a complex trait, and large sample sets are needed to discover multiple genetic factors that underlie it. While in humans such high-throughput studies of IgG N-glycans became usual, only one has been carried out in mice. Here we describe and validate a method for the relative quantification of IgG Fc-linked N-glycans in a subclassspecific manner using nano-reverse phase liquid chromatography coupled with mass-spectrometry (nanoRP-LC-MS) applied to murine IgG. High-throughput data processing is ensured by the LaCyTools software. We have shown that IgG isolation procedure is the main source of technical variation in the current protocol. The major glycoforms were quantified reliably with coefficients of variation below 6% for all the analytes with relative abundances above 5%. We have applied our method to a sample set of 3 inbred strains: BALB/c, C57BL/6 and C3H and observed differences in subclass-specific and strainspecific N-glycosylation of IgG, suggesting a significant genetic component in the regulation of Fclinked IgG N-glycosylation.

Item Type: Article
Uncontrolled Keywords: Protein, N-glycosilation, N-glycan, IgG, nanoRP-LC-MS
Subjects: NATURAL SCIENCES > Biology
Divisions: Division of Molecular Medicine
Depositing User: Ranko Stojković
Date Deposited: 19 Oct 2018 15:50
Last Modified: 19 Oct 2018 15:51
URI: http://fulir.irb.hr/id/eprint/4190
DOI: 10.1038/s41598-018-31844-1

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