title: Study of intact virus-like particles of human papillomavirus by capillary electrophoresis.
abstract: Virus-like particles of human papillomavirus (HPV-VLP), resulting from the self-assembly of the capsid proteins (L1 or L1 and L2), have been widely used to study HPV as they are similar to the native virion. Moreover, two prophylactic vaccines, Gardasil(®) and Cervarix(®), are based on HPV-VLP L1. Analytical techniques currently used to characterize HPV-VLP, such as SDS-PAGE, Western blot, ELISA, are time-consuming and semiquantitative. In this study, CE was evaluated for the analysis of intact HPV16-VLP. The usefulness of capillary inner wall coating as well as various BGEs, pH, and detergent additives were investigated. Reproducible HPV-VLP analysis in CE was achieved using poly(ethylene oxide)-coated capillary and a BGE containing high salt concentration and low SDS concentration. The developed method enables HPV-VLP detection in less than 10 min (migration times RSD: 1.6%). The identity of HPV-VLP peak was confirmed by comparison with a sample obtained from a wild-type baculovirus and with VLP-based vaccine, Gardasil(®) , after adjuvant dissolution. Finally, we applied the developed methodology to VLP-based vaccines, demonstrating that CE could be successfully used for vaccine quality control.
authors.affiliation: Laboratory for the Analysis of Medicines (LAM), Department of Pharmaceutical Sciences, CIRM, University of Liège, Liège, Belgium.
authors.forename: Virginie
authors.initials: V
authors.lastname: Bettonville
authors.affiliation: Cellular and Molecular Immunology, GIGA-Research University of Liège, Liège, Belgium.
authors.forename: Jérôme T J
authors.initials: JT
authors.lastname: Nicol
authors.affiliation: Cellular and Tissular Biology, GIGA-Neurosciences, University of Liège, Liège, Belgium.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Thelen
authors.affiliation: Cellular and Tissular Biology, GIGA-Neurosciences, University of Liège, Liège, Belgium.
authors.forename: Marc
authors.initials: M
authors.lastname: Thiry
authors.affiliation: Laboratory for the Analysis of Medicines (LAM), Department of Pharmaceutical Sciences, CIRM, University of Liège, Liège, Belgium.
authors.forename: Marianne
authors.initials: M
authors.lastname: Fillet
authors.affiliation: Cellular and Molecular Immunology, GIGA-Research University of Liège, Liège, Belgium.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Jacobs
authors.affiliation: Laboratory for the Analysis of Medicines (LAM), Department of Pharmaceutical Sciences, CIRM, University of Liège, Liège, Belgium.
authors.forename: Anne-Catherine
authors.initials: AC
authors.lastname: Servais
mesh_terms: D019075:Electrophoresis, Capillary / Q000379:methods*; D052162:Human papillomavirus 16 / Q000737:chemistry* / Q000302:isolation & purification; D053918:Papillomavirus Vaccines; D011136:Polysorbates; D014771:Virion / Q000737:chemistry* / Q000302:isolation & purification; D014773:Virology / Q000379:methods*
chemical_list: D053918:Papillomavirus Vaccines; D011136:Polysorbates
keywords: Capillary electrophoresis; Human papillomavirus; Surfactant; Virus-like particles