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N-glycosylation analysis of mouse immunoglobulin G isolated from dried blood spots
Authors:Anne-Marie Patenaude  Julija Erhardt  René Hennig  Erdmann Rapp  Gordan Lauc  Marija Pezer
Institution:1. Genos Glycoscience Research Laboratory, Zagreb, Croatia;2. Faculty of Science, University of Zagreb, Zagreb, Croatia;3. glyXera GmbH, Magdeburg, Germany;4. glyXera GmbH, Magdeburg, Germany

Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany;5. Genos Glycoscience Research Laboratory, Zagreb, Croatia

Faculty of Pharmacy and Biochemistry, University of Zagreb, Zagreb, Croatia

Abstract:The association of immunoglobulin G (IgG) glycosylation changes with various human diseases and physiological conditions is well established. Since the mechanistical explanation of the regulation of IgG glycosylation and its functional role in these various states is still missing, the eyes of the biomedical community are now turned towards animal models, which enable intervention studies necessary for conclusions on causality. Mice are recognized and used as a good experimental model for human IgG glycosylation. However, smaller blood volumes, low IgG concentrations at young ages (which are most often used in mice experiments) and multiple sampling protocols during the course of longitudinal studies would profit from a robust workflow for mouse IgG glycome analysis from minute amounts of starting material, collected through a simple sampling procedure. For this purpose, we have developed a protocol for analysis of total N-glycans of IgG isolated from mouse dried blood spots (DBS), which we report here. We show that mouse DBS are a good source of material for IgG N-glycan analysis by multiplexed capillary gel electrophoresis with laser-induced fluorescence (xCGE-LIF).
Keywords:Biomarker  Capillary gel electrophoresis laser-induced fluorescence  Dried blood spots  IgG glycome  Mouse
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