Skip to main content

Easy and Efficient Protocols for Working with Recombinant Vaccinia Virus MVA

  • Protocol
  • First Online:
Vaccinia Virus and Poxvirology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 890))

Abstract

Modified vaccinia virus Ankara (MVA) is a highly attenuated and replication-deficient strain of vaccinia virus that is increasingly used as vector for expression of recombinant genes in the research laboratory and in biomedicine for vaccine development. Major benefits of MVA include the clear safety advantage compared to conventional vaccinia viruses, the longstanding experience in the genetic engineering of the virus, and the availability of established procedures for virus production at an industrial scale. MVA vectors can be handled under biosafety level 1 conditions, and a multitude of recombinant MVA vaccines has proven to be immunogenic and protective when delivering various heterologous antigens in animals and humans. In this chapter we provide convenient state-of-the-art protocols for generation, amplification, and purification of recombinant MVA viruses. Importantly, we include methodology for rigid quality control to obtain best possible vector viruses for further investigations including clinical evaluation.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Moss B (1996) Genetically engineered poxviruses for recombinant gene expression, vaccination, and safety. Proc Natl Acad Sci USA 93:11341–11348

    Article  PubMed  CAS  Google Scholar 

  2. Sutter G, Moss B (1992) Nonreplicating vaccinia vector efficiently expresses recombinant genes. Proc Natl Acad Sci USA 89:10847–10851

    Article  PubMed  CAS  Google Scholar 

  3. Tartaglia J et al (1992) NYVAC: a highly attenuated strain of vaccinia virus. Virology 188:217–232

    PubMed  CAS  Google Scholar 

  4. Acres B, Bonnefoy J-Y (2008) Clinical development of MVA-based therapeutic cancer vaccines. Expert Rev Vaccines 7:889–893

    Article  PubMed  CAS  Google Scholar 

  5. Gomez CE, Najera JL, Krupa M, Esteban M (2008) The poxvirus vectors MVA and NYVAC as gene delivery systems for vaccination against infectious diseases and cancer. Curr Gene Ther 8:97–120

    Article  PubMed  CAS  Google Scholar 

  6. Rimmelzwaan GF, Sutter G (2009) Candidate influenza vaccines based on recombinant modified vaccinia virus Ankara. Expert Rev Vaccines 8:447–454

    Article  PubMed  CAS  Google Scholar 

  7. Kennedy JS, Greenberg RN (2008) IMVAMUNE®: modified vaccinia Ankara strain as an attenuated smallpox vaccine. Expert Rev Vaccines 8:13–24

    Article  Google Scholar 

  8. Mackett M, Smith GL, Moss B (1984) General method for production and selection of infectious vaccinia virus recombinants expressing foreign genes. J Virol 49:857–864

    PubMed  CAS  Google Scholar 

  9. Cottingham MG, Gilbert SC (2010) Rapid generation of markerless recombinant MVA vaccines by en passant recombineering of a self-excising bacterial artificial chromosome. J Virol Methods 168:233–236

    Article  PubMed  CAS  Google Scholar 

  10. Domi A, Moss B (2005) Engineering of a vaccinia virus bacterial artificial chromosome in Escherichia coli by bacteriophage [lambda]-based recombination. Nat Meth 2:95–97

    Article  CAS  Google Scholar 

  11. Wyatt LS et al (2009) Elucidating and minimizing the loss by recombinant vaccinia virus of human immunodeficiency virus gene expression resulting from spontaneous mutations and positive selection. J Virol 83:7176–7184

    Article  PubMed  CAS  Google Scholar 

  12. Meyer H, Sutter G, Mayr A (1991) Mapping of deletions in the genome of the highly attenuated vaccinia virus MVA and their influence on virulence. J Gen Virol 72:1031–1038

    Article  PubMed  CAS  Google Scholar 

  13. Staib C et al (2000) Transient host range selection for genetic engineering of modified vaccinia virus Ankara. Biotechniques 28:1137–1142

    PubMed  CAS  Google Scholar 

  14. Antonis AFG et al (2007) Vaccination with recombinant modified vaccinia virus Ankara expressing bovine respiratory syncytial virus (bRSV) proteins protects calves against RSV challenge. Vaccine 25:4818–4827

    Article  PubMed  CAS  Google Scholar 

  15. Staib C, Lowel M, Erfle V, Sutter G (2003) Improved host range selection for recombinant modified vaccinia virus Ankara. Biotech­niques 34:694–696, 698, 700

    PubMed  CAS  Google Scholar 

  16. Staib C, Drexler I, Sutter G (2004) Construction and isolation of recombinant MVA. Methods Mol Biol 269:77–100

    PubMed  CAS  Google Scholar 

  17. Wong YC, Lin LC, Melo-Silva CR, Smith SA, Tscharke DC (2011) Engineering recombinant poxviruses using a compact GFP-blasticidin resistance fusion gene for selection. J Virol Methods 171:295–8

    Article  PubMed  CAS  Google Scholar 

  18. Renart J, Reiser J, Stark GR (1979) Transfer of proteins from gels to diazobenzyloxymethyl-paper and detection with antisera: a method for studying antibody specificity and antigen structure. Proc Natl Acad Sci USA 76:3116–3120

    Article  PubMed  CAS  Google Scholar 

  19. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by the European Community (FP7 2010; VECTORIE grant No. 261466).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Gerd Sutter .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Kremer, M., Volz, A., Kreijtz, J.H.C.M., Fux, R., Lehmann, M.H., Sutter, G. (2012). Easy and Efficient Protocols for Working with Recombinant Vaccinia Virus MVA. In: Isaacs, S. (eds) Vaccinia Virus and Poxvirology. Methods in Molecular Biology, vol 890. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-876-4_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-876-4_4

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-875-7

  • Online ISBN: 978-1-61779-876-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics