Skip to main content

Production of Chimeric Adenovirus

  • Protocol
  • First Online:
Adenovirus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1089))

  • 2625 Accesses

Abstract

The use of chimeric pseudotyped vectors is a common way to modify the adenoviral tropism by replacing the fiber protein. In this chapter the procedure to generate a chimeric adenovirus pre-stock from a plasmid containing the adenoviral genome is described. Also, the chimeric adenovirus replicative cycle to increase the yield in further productions is determined. Finally, two different protocols, in culture plates and in suspension cultures, to produce the virus at large scale are also detailed.

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 119.00
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. Kojaoghlanian T, Flomenberg P, Horwitz MS (2003) The impact of adenovirus infection on the immunocompromised host. Rev Med Virol 13:155–171

    Article  PubMed  Google Scholar 

  2. Haddada H, Cordier L, Perricaudet M (1995) Gene therapy using adenovirus vectors. Curr Top Microbiol Immunol 199(Pt 3):297–306

    Article  PubMed  CAS  Google Scholar 

  3. Wivel NA, Gao G, Wilson JM (1999) Adenovirus vectors. The development of human gene therapy. Cold Spring Harbor monograph series, pp 87–110

    Google Scholar 

  4. Thomas CE, Ehrhardt A, Kay MA (2003) Progress and problems with the use of viral vectors for gene therapy. Nat Rev Genet 4:346–358

    Article  PubMed  CAS  Google Scholar 

  5. Verma IM, Weitzman MD (2005) Gene therapy: twenty-first century medicine. Annu Rev Biochem 74:711–738

    Article  PubMed  CAS  Google Scholar 

  6. Gall J, Kass-Eisler A, Leinwand L, Falck-Pedersen E (1996) Adenovirus type 5 and 7 capsid chimera: fiber replacement alters receptor tropism without affecting primary immune neutralization epitopes. J Virol 70:2116–2123

    PubMed  CAS  Google Scholar 

  7. Nakamura T, Sato K, Hamada H (2003) Reduction of natural adenovirus tropism to the liver by both ablation of fiber-coxsackievirus and adenovirus receptor interaction and use of replaceable short fiber. J Virol 77:2512–2521

    Article  PubMed  CAS  Google Scholar 

  8. Roberts DM, Nanda A, Havenga MJ, Abbink P, Lynch DM, Ewald BA et al (2006) Hexon-chimaeric adenovirus serotype 5 vectors circumvent pre-existing anti-vector immunity. Nature 441:239–243

    Article  PubMed  CAS  Google Scholar 

  9. Miralles M, Segura MM, Puig M, Bosch A, Chillon M (2012) Efficient amplification of chimeric adenovirus 5/40S vectors carrying the short fiber protein of Ad40 in suspension cell cultures. PLoS One 7:e42073

    Article  PubMed  CAS  Google Scholar 

  10. Sherwood V, Burgert HG, Chen YH, Sanghera S, Katafigiotis S, Randall RE et al (2007) Improved growth of enteric adenovirus type 40 in a modified cell line that can no longer respond to interferon stimulation. J Gen Virol 88:71–76

    Article  PubMed  CAS  Google Scholar 

  11. Tiemessen CT, Kidd AH (1994) Adenovirus type 40 and 41 growth in vitro: host range diversity reflected by differences in patterns of DNA replication. J Virol 68:1239–1244

    PubMed  CAS  Google Scholar 

  12. Rodriguez E, Romero C, Ferrer M, Burgueño JF, Rio A, Gil E, Hamada H, Bosch A, Gasull MA, Fernandez E, Chillon M (2006) Therapeutic potential of the Chimeric Adenovirus 5/40 as a vector for intestine directed gene therapy. Mol Ther 13:S5

    Article  Google Scholar 

  13. Arcasoy SM, Latoche JD, Gondor M, Pitt BR, Pilewski JM (1997) Polycations increase the efficiency of adenovirus-mediated gene transfer to epithelial and endothelial cells in vitro. Gene Ther 4:32–38

    Article  PubMed  CAS  Google Scholar 

  14. Jacobsen F, Hirsch T, Mittler D, Schulte M, Lehnhardt M, Druecke D et al (2006) Polybrene improves transfection efficacy of recombinant replication-deficient adenovirus in cutaneous cells and burned skin. J Gene Med 8:138–146

    Article  PubMed  CAS  Google Scholar 

  15. Geisse S, Di Maiuta N, Ten Buren B, Henke M (2005) The secrets of transfection in serum-free suspension culture. In: Gòdia F, Fussenegger M (eds) Animal cell technology meets genomics. ESACT Proceedings. Springer, Netherlands, pp 373–376

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Miralles, M., Garcia, M., Tejero, M., Bosch, A., Chillón, M. (2014). Production of Chimeric Adenovirus. In: Chillón, M., Bosch, A. (eds) Adenovirus. Methods in Molecular Biology, vol 1089. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-679-5_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-679-5_16

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-678-8

  • Online ISBN: 978-1-62703-679-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics