Skip to main content

Simulated Digestion for Testing the Stability of Edible Vaccine Based on Cucumber mosaic virus (CMV) Chimeric Particle Display Hepatitis C virus (HCV) Peptide

  • Protocol
  • First Online:
Virus Hybrids as Nanomaterials

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1108))

Abstract

Edible vaccines must survive digestive process and preserve the specific structure of the antigenic peptide to elicit effective immune response. The stability of a protein to digestive process can be predicted by subjecting it to the in vitro assay with simulated gastric fluid (SGF) and simulated intestinal fluid (SIF). Here, we describe the protocol of producing and using chimeric Cucumber mosaic virus (CMV) displaying Hepatitis C virus (HCV) derived peptide (R9) in double copy as an oral vaccine. Its stability after treatment with SGF and SIF and the preservation of the antigenic properties were verified by SDS-PAGE and immuno western blot techniques.

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 109.99
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. Takagi K, Teshima R, Okunuki H, Sawada J (2003) Comparative study of in vitro digestibility of food protein and effect of preheating on the digestion. Biol Pharm Bull 26(7): 969–973

    Article  CAS  Google Scholar 

  2. Huang YY, Liu GM, Cai QF et al (2010) Stability of major allergen tropomyosin and other food proteins of mud crab (Scylla serrata) by in vitro gastrointestinal digestion. Food Chemical Toxicol 48:1196–1201

    Article  CAS  Google Scholar 

  3. Rae CS, Khor IW, Wang Q et al (2005) Systemic trafficking of plant virus nanoparticles in mice via the oral route. Virology 343:224–235

    Article  CAS  Google Scholar 

  4. Nuzzaci M, Vitti A, Condelli V et al (2010) In vitro stability of Cucumber mosaic virus nanoparticles carrying a Hepatitis C virus-derived epitope under simulated gastrointestinal conditions and in vivo efficacy of an edible vaccine. J Virol Methods 165:211–215

    Article  CAS  Google Scholar 

  5. Taylor SL (2002) Protein allergenicity assessment of foods produced through agricultural biotechnology. Annu Rev Pharmacol Toxicol 42:99–112

    Article  CAS  Google Scholar 

  6. Joensuu JJ, Kotiaho M, Riipi T et al (2004) Fimbrial subunit protein FaeG expressed in transgenic tobacco inhibits the binding of F4ac enterotoxigenic Escherichia coli to porcine enterocytes. Transgenic Res 13:295–298

    Article  CAS  Google Scholar 

  7. Des Rieux A, Fievez V, Garinot M, Schneider YJ, Preat V (2006) Nanoparticles as potential oral delivery systems of protein and vaccines: a mechanistic approach. J Control Rel 116: 1–27

    Article  CAS  Google Scholar 

  8. Puntoriero G, Meola A, Lahm A et al (1998) Towards a solution for Hepatitis C virus hypervariability: mimotopes of the hypervariable region 1 can induce antibodies cross-reacting with a large number of viral variants. EMBO J 17(13):3521–3533

    Article  CAS  Google Scholar 

  9. Nuzzaci M, Piazzolla G, Vitti A et al (2007) Cucumber mosaic virus as a presentation system for a double hepatitis C virus-derived epitope. Arch Virol 152:915–928

    Article  CAS  Google Scholar 

  10. Lot H, Marrou J, Quiot JB, Esvan C (1972) Contribution a l’etude du virus de la mosaique du cocombre (CMV). I. Methode de purification rapide du virus. Ann Phytopathol 14: 25–38

    Google Scholar 

  11. Natilla A, Piazzolla G, Nuzzaci M et al (2004) Cucumber mosaic virus as carrier of a Hepatitis C virus-derived epitope. Arch Virol 149(1): 137–154

    Article  CAS  Google Scholar 

  12. Vitti A, Piazzolla G, Condelli V et al (2010) Cucumber mosaic virus as the expression system for a potential vaccine against Alzheimer’s disease. J Virol Methods 69:332–340

    Article  Google Scholar 

  13. Feng JL, Chen SN, Tang XS et al (2006) Quantitative determination of cucumber mosaic virus genome RNAs in virions by realtime transcription-polymerase chain reaction. Acta Biochim Biophys Sin 38:669–676

    Article  CAS  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, New York

About this protocol

Cite this protocol

Vitti, A., Nuzzaci, M., Condelli, V., Piazzolla, P. (2014). Simulated Digestion for Testing the Stability of Edible Vaccine Based on Cucumber mosaic virus (CMV) Chimeric Particle Display Hepatitis C virus (HCV) Peptide. In: Lin, B., Ratna, B. (eds) Virus Hybrids as Nanomaterials. Methods in Molecular Biology, vol 1108. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-751-8_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-751-8_3

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-750-1

  • Online ISBN: 978-1-62703-751-8

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics