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Archives of Virology

, Volume 153, Issue 3, pp 585–589 | Cite as

An investigation into the use of human papillomavirus type 16 virus-like particles as a delivery vector system for foreign proteins: N- and C-terminal fusion of GFP to the L1 and L2 capsid proteins

  • Oliver P. Windram
  • Brandon Weber
  • Mohamed A. Jaffer
  • Edward P. Rybicki
  • Dionne N. Shepherd
  • Arvind Varsani
Brief Report

Abstract

Development of vaccine strategies against human papillomavirus (HPV), which causes cervical cancer, is a priority. We investigated the use of virus-like particles (VLPs) of the most prevalent type, HPV-16, as carriers of foreign proteins. Green fluorescent protein (GFP) was fused to the N or C terminus of both L1 and L2, with L2 chimeras being co-expressed with native L1. Purified chimaeric VLPs were comparable in size (∼55 nm) to native HPV VLPs. Conformation-specific monoclonal antibodies (Mabs) bound to the VLPs, thereby indicating that they possibly retain their antigenicity. In addition, all of the VLPs encapsidated DNA in the range of 6–8 kb.

Keywords

Green Fluorescent Protein Capsid Protein Fusion Construct Green Fluorescent Protein Fusion Capsid Shell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We would like thank to Dr. Neil Christensen for providing the monoclonal antibodies used in this study. This research was funded by the Poliomyelitis Research Foundation of South Africa (grant #04/25) and the University of Cape Town start up grant. AV is supported by the Carnegie Corporation of New York.

Supplementary material

705_2007_25_MOESM1_ESM.doc (37 kb)
Supplementary Table 1: Primer sets used for cloning fusion constructs (DOC 37 kb)
705_2007_25_MOESM2_ESM.doc (834 kb)
Supplementary Figure 1: CsCl banding patterns observed for the fusion constructs. Two bands observed for GFP-L1 and L1-GFP constructs with the lower band fluorescing under UV (GFP). Three bands were observed for L1:GFP-L1 and L1:L2-GFP. (DOC 834 kb)
705_2007_25_MOESM3_ESM.doc (114 kb)
Supplementary Figure 2: Agarose gel stained with ethidium bromide, showing the banding of 6-8-kb DNA incorporated into the GFP fusion VLPs. A greater amount of DNA with a broader size range (6-8kb) was observed for GFP-L1 and L1-GFP. DNA of smaller size was observed in the L1:GFP-L2 and L1:L2-GFP VLPs. The fluorescence in the wells represents the GFP fusion VLPs. (DOC 114 kb)

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Oliver P. Windram
    • 1
  • Brandon Weber
    • 2
  • Mohamed A. Jaffer
    • 2
  • Edward P. Rybicki
    • 1
  • Dionne N. Shepherd
    • 1
  • Arvind Varsani
    • 1
    • 2
  1. 1.Department of Molecular and Cell BiologyUniversity of Cape TownCape TownSouth Africa
  2. 2.Electron Microscope UnitUniversity of Cape TownCape TownSouth Africa

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