Vaccine Design pp 155-166 | Cite as

Development of Rabies Virus-Like Particles for Vaccine Applications: Production, Characterization, and Protection Studies

  • Diego Fontana
  • Marina Etcheverrigaray
  • Ricardo Kratje
  • Claudio Prieto
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1403)

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.

Key words

Vaccine development Virus-like particles Rabies Lentiviral vectors Stable cell line HEK293 

Notes

Acknowledgements

The authors would like to thank Juan Pablo Soto (Institutional Image, Universidad Nacional del Litoral, Argentina) for technical support.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Diego Fontana
    • 1
    • 2
  • Marina Etcheverrigaray
    • 1
  • Ricardo Kratje
    • 1
  • Claudio Prieto
    • 1
    • 2
  1. 1.Cell Culture Laboratory, Biochemistry and Biological Sciences SchoolUniversidad Nacional del LitoralSanta FeArgentina
  2. 2.Biotechnological Development Laboratory, Biochemistry and Biological Sciences SchoolUniversidad Nacional del LitoralSanta FeArgentina

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