Abstract
The ongoing threat of pathogens, increasing resistance against antibiotics, and the risk of fast spreading of infectious diseases in a global community resulted in an intensified development of vaccines. Antigens used for vaccination comprise a wide variety of macromolecules including glycoproteins, lipopolysaccharides, and complex carbohydrates. For all of these antigens the sugar composition plays a crucial role for immunogenicity and protective efficacy of the vaccine. Here, we provide a protocol for N-glycosylation fingerprinting utilizing high performance multiplexed capillary gel electrophoresis with laser-induced fluorescence detection (xCGE-LIF) technology. The method described, enables to analyze the N-glycosylation of specific proteins out of a complex sample or even the total of all N-glycans contained in such a sample. The protocol is exemplarily demonstrated for N-glycosylation fingerprinting of cell culture-derived influenza A and B viruses and their major antigens, the membrane glycoproteins hemagglutinin and neuraminidase.
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*Both authors contributed equally
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Acknowledgements
This work was supported by the Max Planck Society and by the European Union’s Seventh Framework Programme (FP7-Health-F5-2011) under grant agreement no. 278535 “HighGlycan.”
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Hennig, R., Rapp, E., Kottler, R., Cajic, S., Borowiak, M., Reichl, U. (2015). N-Glycosylation Fingerprinting of Viral Glycoproteins by xCGE-LIF. In: Lepenies, B. (eds) Carbohydrate-Based Vaccines. Methods in Molecular Biology, vol 1331. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2874-3_8
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DOI: https://doi.org/10.1007/978-1-4939-2874-3_8
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