N-Glycosylation Fingerprinting of Viral Glycoproteins by xCGE-LIF.

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.

DOI: 10.1007/978-1-4939-2874-3_8

Cite this paper

@article{Hennig2015NGlycosylationFO, title={N-Glycosylation Fingerprinting of Viral Glycoproteins by xCGE-LIF.}, author={Ren{\'e} Hennig and Erdmann Rapp and Robert Kottler and Samanta Cajic and Matthias Borowiak and Udo Reichl}, journal={Methods in molecular biology}, year={2015}, volume={1331}, pages={123-43} }