Abstract
The ability to make a large variety of virus-like particles (VLPs) has been successfully achieved in the baculovirus expression vector system (BEVS)/insect cell system. The production and scale-up of these particles, which are mostly sought as candidate vaccines, are currently being addressed. Furthermore, these VLPs are being investigated as delivery agents for use as therapeutics. Recently, adeno-associated viral (AAV) vectors, which can be potentially used for human gene therapy, have been produced in insect cells using three baculovirus vectors to supply the required genes. The use of host insect cells allows mass production of VLPs in a proven scaleable system. This chapter focuses on the methodology, based on the work done in our lab, for the production of AAV-like particles and vectors in a BEVS/insect cell system.
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References
Zheng, Y. Z., Greenfield, P. F., and Reid, S. (1999) Optimized production of recombinant bluetongue core-like particles produced by the baculovirus expression system. Biotechno. Bioeng. 65, 600–604.
Maranga, L., Cruz, P. E., Aunins, J., and Carrondo, M. J. T. (2002) Production of core and virus-like particles with baculovirus infected cells. Adv. Biochem. Eng. Biotechnol. 74, 183–206.
Palomares, L. A., Lopez, S., and Ramirez, O. T. (1999) in Animal Cell Technology: Products from Cells, Cells as Products (Bernard, A., ed.), Kluwer Academic Publishers, Boston, pp. 347–349.
Palomares, L. A., Lopez, S., and Ramirez, O. T. (2002) Strategies for manipulating the relative concentration of recombinant rotavirus structural proteins during simultaneous production by insect cells. Biotechnol. Bioeng. 78, 635–644.
Jiang, B., Barniak, V., Smith, R. P., et al. (1998) Synthesis of rotavirus-like particles in insect cells: comparative and quantitative analysis. Biotechnol. Bioeng. 60, 369–374.
Park, J. Y., Kim, H., Hwang, H. K., et al. (2004) Large-scale production of rotavirus VLP as vaccine candidate using baculovirus expression vector system (BEVS). J. Microbiol. Biotechnol. 14, 35–40.
Tsao, E. I., Mason, M. R., Cacciuttolo, M. A., Bowen, S. H., and Folena-Wasserman, G. (1996) Production of parvovirus B19 vaccine in insect cells co-infected with double baculoviruses. Biotechnol. Bioeng. 49, 130–138.
Maranga, L., Brazao, T. F., and Carrondo, M. J. T. (2003) Virus-like particle production at low multiplicities of infection with the baculovirus insect cell system. Biotechnol. Bioeng. 84, 245–253.
Maranga, L., Cunha, A., Clemente, J., Cruz, P., and Carrondo, M. J. T. (2004) Scale-up of virus-like particles production: effects of sparging, agitation and bioreactor scale on cell growth, infection kinetics and productivity. J. Biotechnol. 107, 55–64.
Maranga, L., Rueda, P., Antonis, A. F. G., et al. (2002) Large scale production and downstream processing of a recombinant porcine parvovirus vaccine. Appl. Microbiol. Biotechnol. 59, 45–50.
Cruz, P. E., Maranga, L., and Carrondo, M. J. T. (2002) Integrated process optimization: lessons from retrovirus and virus-like particle production. J. Biotechnol. 99, 199–214.
Cruz, P. E., Cunha, A., Peixoto, C. C., Clemente, J., Moreira, J. L., and Carrondo, M. J. T. (1998) Optimization of the production of virus-like particles in insect cells. Biotechnol. and Bioeng. 60, 408–418.
Cruz, P. E., Martins, P. C., Alves, P. M., et al. (1999) Proteolytic activity in infected and noninfected insect cells: degradation of HIV-1 Pr55gag particles. Biotechnol. Bioeng. 65, 133–143.
Hu, Y.-C. and Bentley, W. E. (2000) A kinetic and statistical-thermodynamic model for baculovirus infection and virus-like particle assembly in suspended insect cells. Chem. Eng. Sci. 55, 3991–4008.
Hu, Y.-C. and Bentley, W. E. (2001) Effect of MOI ratio on the composition and yield of chimeric infectious bursal disease virus-like particles by baculovirus co-infection: deterministic predictions and experimental results. Biotechnol. Bioeng. 75, 104–119.
Hu, Y.-C., Bentley, W. E., Edwards, G. H., and Vakharia, V. (1999) Chimeric infectious bursal disease virus-like particles expressed in insect cells and purified by immobilized metal affinity chromatography. Biotechnol. Bioeng. 63, 721–729.
Urabe, M., Ding, C., and Kotin, R. M. (2002) Insect cells as a factory to produce adeno-associated virus type 2 vectors. Hum. Gene Ther. 13, 1935–1943.
Meghrous, J., Aucoin, M. G., Jacob, D., Chahal, P. S., Arcand, N., and Kamen, A. A. (2005) Production of recombinant adeno-associated viral vectors using a baculovirus/insect cell suspension culture system: from shake flasks to a 20-L bioreactor. Biotechnol. Prog. 21, 154–160.
Hoque, M., Shimizu, N., Ishizu, K.-I., et al. (1999) Chimeric virus-like particle formation of adeno-associated virus. Biochem. Biophys. Res. Comm. 266, 371–376.
Ruffing, M., Zentgraf, H., and Kleinschmidt, J. A. (1992) Assembly of viruslike particles by recombinant structural proteins of adeno-associated virus type 2 in insect cells. J. Virol. 66, 6922–6930.
Shen, C. F., Meghrous, J., and Kamen, A. (2002) Quantitation of baculovirus particles by flow cytometry. J. Virol. Meth. 105, 321–330.
Zeiser, A., Elias, C. B., Voyer, R., Jardin, B., and Kamen, A. (2000) On-line monitoring of physiological parameters of insect cell cultures during the growth and infection process. Biotechnol. Prog. 16, 803–808.
Zolotukhin, S., Byrne, B. J., Mason, E., et al. (1999) Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther. 6, 973–985.
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Aucoin, M.G., Jacob, D., Chahal, P.S., Meghrous, J., Bernier, A., Kamen, A.A. (2007). Virus-like Particle and Viral Vector Production Using the Baculovirus Expression Vector System/Insect Cell System. In: Murhammer, D.W. (eds) Baculovirus and Insect Cell Expression Protocols. Methods in Molecular Biology™, vol 388. Humana Press. https://doi.org/10.1007/978-1-59745-457-5_14
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DOI: https://doi.org/10.1007/978-1-59745-457-5_14
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