title: Characterization of N-glycosylation profiles from mammalian and insect cell derived chikungunya VLP.
abstract: Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes severe arthralgia. The envelope of CHIKV is composed of 240 copies of two glycoproteins: E1 and E2. In this work, we have characterized the N-glycosylation patterns of CHIKV virus-like particles (VLPs), containing both E1 and E2 proteins, derived from mammalian and insect cells using hydrophilic interaction liquid chromatography (HILIC) with fluorescence (FL) and mass spectrometry (MS) detection. While HEK293 derived CHIKV VLPs contain oligomannose, hybrid and complex glycans, VLPs derived from SfBasic predominantly contain oligomannose glycans. This strong host dependence of N-glycosylation pattern resembles other alphaviruses such as SINV. The VLPs from HEK293 and SfBasic, with significantly different N-glycosylation profiles, are valuable reagents enabling future in-depth correlation studies between immunogenicity and glycosylation. In addition, the characterization tools presented here allow one to monitor glycosylation during vaccine process development and ensure process consistency.
authors.affiliation: Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States.
authors.forename: Catherine
authors.initials: C
authors.lastname: Lancaster
authors.affiliation: Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States.
authors.forename: Pavlo
authors.initials: P
authors.lastname: Pristatsky
authors.affiliation: Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States.
authors.forename: Van M
authors.initials: VM
authors.lastname: Hoang
authors.affiliation: Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States.
authors.forename: Danilo R
authors.initials: DR
authors.lastname: Casimiro
authors.affiliation: Vaccine Production Program, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Gaithersburg, MD 20878, United States.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Schwartz
authors.affiliation: Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States.
authors.forename: Richard
authors.initials: R
authors.lastname: Rustandi
authors.affiliation: Vaccine Bioprocess Research & Development, Merck Research Laboratories, West Point, PA, 19486, United States. Electronic address: sha_ha@merck.com.
authors.forename: Sha
authors.initials: S
authors.lastname: Ha
mesh_terms: D000818:Animals; D002460:Cell Line; D065632:Chikungunya Fever / Q000821:virology*; D002646:Chikungunya virus / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000379:methods; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007313:Insecta; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D011134:Polysaccharides / Q000032:analysis*; D014759:Viral Envelope Proteins / Q000737:chemistry*
chemical_list: D011134:Polysaccharides; D014759:Viral Envelope Proteins
keywords: Chikungunya; Glycosylation; HILIC; VLP; Vaccine