engleski

Dried blood spots as source material for immunoglobulin G N-glycan analysis

Aim: Immunoglobulin G (IgG) N-glycans are emerging as a new putative biomarker for different diseases and are routinely released from IgG isolated from plasma or serum samples. Since dried blood spots (DBS) have many advantages over plasma or serum samples (reduction of sample volume, simplification of the collection, storage and transport procedures, decreased cost of analysis), the aim of this study was to evaluate DBS as source material for IgG N-glycan analysis. Material and methods: IgG was isolated from DBS by affinity chromatography on a protein G monolithic plate and its N glycans released by PNGase F, labeled with 2 aminobenzamide and analyzed by hydrophilic interaction ultraperformance liquid chromatography (UPLC). Results: IgG isolation from DBS and N glycan analysis by UPLC were very robust and the N glycan profile obtained from IgG isolated from DBS was comparable to the profile obtained from IgG isolated from plasma. The IgG N-glycan profiles obtained from DBS stored at -20°C with desiccant remained stable for up to 29 days. Conclusion: DBS are a good and stable source material for a robust IgG N-glycan analysis by UPLC, thus enabling a comprehensive IgG total N-glycan analysis even in cases where no trained personnel and/or laboratory are available for sample collection.

biomarker ; dried blood spots ; high-throughput ; IgG glycome ; ultra-performance liquid chromatography

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