Skip to main content
Log in

Adenovirus Dodecahedron, a VLP, Can be Purified by Size Exclusion Chromatography Instead of Time-Consuming Sucrose Density Gradient Centrifugation

  • Research
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

Adenoviral dodecahedron (Dd) is a virus-like particle composed of twelve pentameric penton base (Pb) proteins, responsible for adenovirus cell penetration. It is generated spontaneously in the baculovirus system upon expression of the Pb gene of adenovirus serotype 3. This particle shows remarkable cell penetration ability with 2,00,000–3,00,000 Dd internalized into one cell in culture, conceivably delivering several millions of foreign cargo molecules to the target cell. We have used it in the past for delivery of small drugs as well as a vaccination platform, in which Dd serves as a particulate vaccine delivery system. Since development of new biomedicals depends strongly on the cost of their expression and purification, we attempted, albeit unsuccessfully, to obtain Dd expression in bacteria. We therefore retained its expression in the baculovirus/insect cells system but introduced significant improvements in the protocols for Dd expression and purification, leading to considerable savings in time and improved yield.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

Ad:

Adenovirus

Ad3:

Adenovirus serotype 3

BSA:

Bovine serum albumin

CBB:

Coomassie Brilliant Blue

Dd:

Dodecahedron

EM:

Electron microscopy

FBS:

Fetal bovine serum

FT:

Flow-through

HRP:

Horseradish peroxidase

IEC:

Ion exchange chromatography

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

MW:

Molecular weight

Pb:

Penton base protein

PBS:

Phosphate buffered saline

SDS-PAGE:

SDS polyacrylamide gel electrophoresis

SEC:

Size exclusion chromatography

TMB:

3,3′,5,5′-Tetramethylbenzidine

VLP:

Virus-like particle

WB:

Western blot

References

  1. Wickham, T. J., Mathias, P., Cheresh, D. A., & Nemerow, G. R. (1993). Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment. Cell, 73(2), 309–319.

    Article  CAS  Google Scholar 

  2. Fender, P., Ruigrok, R. W., Gout, E., Buffet, S., & Chroboczek, J. (1997). Adenovirus dodecahedron, a new vector for human gene transfer. Nature Biotechnology, 15(1), 52–56.

    Article  CAS  Google Scholar 

  3. Fender, P., Schoehn, G., Foucaud-Gamen, J., Gout, E., Garcel, A., Drouet, E., & Chroboczek, J. (2003). Adenovirus dodecahedron allows large multimeric protein in human cells. Journal of Virology, 77(8), 4960–4964.

    Article  CAS  Google Scholar 

  4. Garcel, A., Gout, E., Timmins, J., Chroboczek, J., & Fender, P. (2006). Protein transduction into human cells by adenovirus dodecahedron using WW domains as universal adaptors. Journal of Gene Medicine, 8(4), 524–531.

    Article  CAS  Google Scholar 

  5. Szolajska, E., Burmeister, W. P., Zochowska, M., Nerlo, B., Andreev, I., Schoehn, G., et al. (2012). The structural basis for the integrity of adenovirus Ad3 dodecahedron. PLoS One, 7(9), e46075.

    Article  CAS  Google Scholar 

  6. Zochowska, M., Paca, A., Schoehn, G., Andrieu, J. P., Chroboczek, J., Dublet, B., & Szolajska, E. (2009). Adenovirus dodecahedron, as a drug delivery vector. PLoS One, 4, e5569.

    Article  Google Scholar 

  7. Zochowska, M., Piguet, A. C., Jemielity, J., Kowalska, J., Szolajska, E., Dufour, J. F., & Chroboczek, J. (2014). Virus-like particle-mediated intracellular delivery of mRNA cap analog with in vivo activity against hepatocellular carcinoma. Nanomedicine, S1549–9634(14), 00419.

    Google Scholar 

  8. Szurgot, I., Szolajska, E., Laurin, D., Lambrecht, B., Chaperot, L., Schoehn, G., & Chroboczek, J. (2013). Self-adjuvanting influenza candidate vaccine presenting epitopes for cell-mediated immunity on a proteinaceous multivalent nanoplatform. Vaccine, 31(40), 4338–4346.

    Article  CAS  Google Scholar 

  9. Gallegos, C. O., & Patton, J. T. (1989). Characterization of rotavirus replication intermediates: A model for the assembly of single-shelled particles. Virology, 172(2), 616–627.

    Article  CAS  Google Scholar 

  10. Hitchman, R. B., Possee, R. D., Siaterli, E., Richards, K. S., Clayton, A. J., Bird, L. E., et al. (2010). Improved expression of secreted and membrane-targeted proteins in insect cells. Biotechnology and Applied Biochemistry, 56(3), 85–93.

    Article  CAS  Google Scholar 

  11. Urbas, L., Lah Jarc, B., Barut, M., Zochowska, M., Chroboczek, J., Pihlar, B., & Szolajska, E. (2011). Purification of recombinant adenovirus type 3 dodecahedric virus-like particles for biomedical applications using short monolithic columns. Journal of Chromatography A, 1218(17), 2451–2459.

    Article  CAS  Google Scholar 

  12. Naskalska, A., Szolajska, E., Chaperot, L., Angel, J., Plumas, J., & Chroboczek, J. (2009). Influenza recombinant vaccine: Matrix protein M1 on the platform of the adenovirus dodecahedron. Vaccine, 27(52), 7385–7393.

    Article  CAS  Google Scholar 

  13. Savard, C., Laliberté-Gagné, M. È., Babin, C., Bolduc, M., Guérin, A., Drouin, K., et al. (2012). Improvement of the PapMV nanoparticle adjuvant property through an increase of its avidity for the antigen [influenza NP]. Vaccine, 30(15), 2535–2542.

    Article  CAS  Google Scholar 

  14. Teunissen, E. A., de Raad, M., & Mastrobattista, E. (2013). Production and biomedical applications of virus-like particles derived from polyomaviruses. Journal of Controlled Release, 172(1), 305–321.

    Article  CAS  Google Scholar 

  15. Wei, M. X., Zhang, X., Yu, H., Tang, Z. M., Wang, K. H., Li, Z. Y., et al. (2014). Bacteria expressed hepatitis E virus capsid proteins maintain virion-like epitopes. Vaccine, 32(24), 2859–2865.

    Article  CAS  Google Scholar 

  16. Chuan, Y. P., Fan, Y. Y., Lua, L. H. L., & Middelberg, A. P. J. (2010). Virus assembly occurs following a pH- or Ca2+-triggered switch in the thermodynamic attraction between structural protein capsomeres. Journal of the Royal Society, Interface, 7(44), 409–421.

    Article  CAS  Google Scholar 

  17. Liew, M. W. O., Chuan, Y. P., & Middelberg, A. P. J. (2012). High-yield and scalable cellfree assembly of virus-like particles by dilution. Biochemical Engineering Journal, 67, 88–96.

    Article  CAS  Google Scholar 

  18. Simon, C., Klose, T., Herbst, S., Han, B. G., Sinz, A., Glaeser, R. M., et al. (2014). Disulfide linkage and structure of highly stable yeast-derived virus-like particles of murine polyomavirus. Journal of Biological Chemistry, 289(15), 10411–10418.

    Article  CAS  Google Scholar 

  19. Fender, P., Boussaid, A., Mezin, P., & Chroboczek, J. (2005). Synthesis, cellular localization, and quantification of penton-dodecahedron in serotype 3 adenovirus-infected cells. Virology, 340(2), 167–173.

    Article  CAS  Google Scholar 

  20. Vicente, T., Roldão, A., Peixoto, C., Carrondo, M. J., & Alves, P. M. (2011). Large-scale production and purification of VLP-based vaccines. Journal of Invertebrate Pathology, 107(Suppl), S42–488. Review.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by Polish National Science Center (DEC-2013/09/B/NZ3/02327). The electron microscope facility used in this work is supported by the Rhône-Alpes Region, FRM, FEDER of CNRS, CEA, University of Grenoble, EMBL, and the GIS-Infrastructures of IBISA. We are grateful to Irina Kikhno (Institute of Molecular Biology & Genetics of Ukrainian Academy of Science, Kiev, Ukraine) for the plasmid for construction of the E. coli expression vector and to Elzbieta Grzesiuk’s team for their collaboration in protein purification.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to J. Chroboczek.

Additional information

I. Szurgot and M. Jedynak contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Szurgot, I., Jedynak, M., Podsiadla-Bialoskorska, M. et al. Adenovirus Dodecahedron, a VLP, Can be Purified by Size Exclusion Chromatography Instead of Time-Consuming Sucrose Density Gradient Centrifugation. Mol Biotechnol 57, 565–573 (2015). https://doi.org/10.1007/s12033-015-9850-9

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12033-015-9850-9

Keywords

Navigation