title: Development of Rabies Virus-Like Particles for Vaccine Applications: Production, Characterization, and Protection Studies.
abstract: Rabies is a viral infection of the central nervous system for which vaccination is the only treatment possible. Besides preexposure, vaccination is highly recommended for people living in endemic areas, veterinarians, and laboratory workers. Our group has developed rabies virus-like particles (RV-VLPs) with immunogenic features expressed in mammalian cells for vaccine applications. In this chapter the methods to obtain and characterize a stable HEK293 cell line expressing RV-VLPs are detailed. Further, analytical ultracentrifugation steps to purify the obtained VLPs are developed, as well as western blot, dynamic light scattering, and immunogold electron microscopy to analyze the size, distribution, shape, and antigenic conformation of the purified particles. Finally, immunization protocols are described to study the immunogenicity of RV-VLPs.
authors.affiliation: Cell Culture Laboratory, Biochemistry and Biological Sciences School, Universidad Nacional del Litoral, Bv. Pellegrini 2750, Santa Fe, 3000, Argentina. diego_sf@hotmail.com.|Biotechnological Development Laboratory, Biochemistry and Biological Sciences School, Universidad Nacional del Litoral, Santa Fe, Argentina. diego_sf@hotmail.com.
authors.forename: Diego
authors.initials: D
authors.lastname: Fontana
authors.affiliation: Cell Culture Laboratory, Biochemistry and Biological Sciences School, Universidad Nacional del Litoral, Bv. Pellegrini 2750, Santa Fe, 3000, Argentina.
authors.forename: Marina
authors.initials: M
authors.lastname: Etcheverrigaray
authors.affiliation: Cell Culture Laboratory, Biochemistry and Biological Sciences School, Universidad Nacional del Litoral, Bv. Pellegrini 2750, Santa Fe, 3000, Argentina.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Kratje
authors.affiliation: Cell Culture Laboratory, Biochemistry and Biological Sciences School, Universidad Nacional del Litoral, Bv. Pellegrini 2750, Santa Fe, 3000, Argentina.|Biotechnological Development Laboratory, Biochemistry and Biological Sciences School, Universidad Nacional del Litoral, Santa Fe, Argentina.
authors.forename: Claudio
authors.initials: C
authors.lastname: Prieto
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D011820:Rabies virus / Q000276:immunology*; D058425:Vaccines, Virus-Like Particle / Q000276:immunology* / Q000302:isolation & purification; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D058425:Vaccines, Virus-Like Particle; D014765:Viral Vaccines
keywords: HEK293; Lentiviral vectors; Rabies; Stable cell line; Vaccine development; Virus-like particles