title: 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.
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstrasse1, 39106, Magdeburg, Germany.
authors.forename: René
authors.initials: R
authors.lastname: Hennig
authors.forename: Erdmann
authors.initials: E
authors.lastname: Rapp
authors.forename: Robert
authors.initials: R
authors.lastname: Kottler
authors.forename: Samanta
authors.initials: S
authors.lastname: Cajic
authors.forename: Matthias
authors.initials: M
authors.lastname: Borowiak
authors.forename: Udo
authors.initials: U
authors.lastname: Reichl
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000737:chemistry* / Q000276:immunology; D018929:Cell Culture Techniques; D004285:Dogs; D019075:Electrophoresis, Capillary / Q000379:methods; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology; D006031:Glycosylation; D006388:Hemagglutinins / Q000737:chemistry / Q000276:immunology; D009980:Influenza A virus / Q000276:immunology; D009981:Influenza B virus / Q000276:immunology; D007252:Influenza Vaccines / Q000276:immunology; D061985:Madin Darby Canine Kidney Cells; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000276:immunology; D009439:Neuraminidase / Q000737:chemistry / Q000276:immunology; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D000956:Antigens, Viral; D006023:Glycoproteins; D006388:Hemagglutinins; D007252:Influenza Vaccines; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D014764:Viral Proteins; D009439:Neuraminidase