Abstract
Low cell density, suboptimal nutrient environment and increased down time, and labour of repeated batch fermentations impose major limitations to high product yields in recombinant baculovirus infected insect cell cultures.
In an attempt to reduce these limitations for the production of heterologous protein (γ-galactosidase) and virus (rAcNPV) by the Baculovirus Expression Vector System employing a Spodoptera frugiperda (Sf-9) cell line, a two-stage bioreactor system was developed. The first stage bioreactor was designed for the continuous cultivation of insect cells while the second-stage bioreactor was intended for continuous viral infection of the cells at high density.
The system allowed continuous high cell density infection while maintaining optimal specific recombinant virus and protein yields as determined by a batch control. The extended run-time and increased specific production of the system resulted in enhanced volumetric production compared with batch, before the yields declined apparently due to the dominance of defective interfering particles (DIPS).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Caron A.W., Archambault J., Massie B. (1990) High level recombinant protein production in bioreactors using the baculovirus-insect cell expression system. Biotechnology and Bioengineering, 36: 1133–1140.
Cavegn C. and Bernard, A.R. (1992) A Perfusion Process For High Density Insect Cell Cultures. In: Baculovirus and Recombinant Protein Production Processes. Eds. J.M. Vlak, E.J. Schlaeger and A.R. Bernard, Editiones Roche, Basel, Switzerland, 262–273.
de Gooijer, C.D., van Lier, F.L.J., van den End, E.J., Vlak, J.M., and Tramper, J. (1989) A model for Baculovirus production with continuous insect cell cultures. Appl Microbiol Biotechnol 30:497–501.
Jäger, V., Grabenhorst, E., Kobold, A., Deutschmann, S., and Conradt, H.S. (1992) High Density Perfusion Culture of Insect Cells for the Production of Recombinant Glycoproteins. In: Baculovirus and Recombinant Protein Production Processes. Eds. J.M. Vlak, E.J. Schlaeger and A.R. Bernard, Editiones Roche, Basel, Switzerland, 274–288.
Kompier R., Tramper J., Vlak J.M. (1988) A continuous process for the production of baculovirus using insect-cell cultures. Biotechnology Letters, 10(12): 849–854.
van Lier, F.L.J., van den End E.J., de Gooijer CD., Vlak J.M., Tramper J. (1990) Continuous production of baculoviruses in a cascade of insect-cell bioreactors. Applied Microbiology and Biotechnology, 33: 43–47.
van Lier, F.L.J., Vlak J.M., Tramper J. (1992) Production of baculovirus-expressed proteins from suspension cultures of insect cells. Animal Cell Biotechnology, 5: 169–188.
Nielsen L.K., Smyth G.K., Greenfield P.F. (1991) Haemocytometer cell count distributions: implications of non-poisson behaviour. Biotechnology Progress, 7: 560–563.
Nielsen L.K., Smyth G.K., Greenfield P.F. (1992) Accuracy of the endpoint assay for virus titration. Cytotechnology, 8: 231–236.
Radford K.R., Reid S., and Greenfield P.F. (1991) Improved production of recombinant proteins by the baculovirus expression vector system. In: Animal Cell Technology-Basic and Applied Aspects. H. Murakami, S. Shirahata, and H. Tachibana (Eds). Kluwer Academic Publishers, Netherlands, 391–396.
Radford K.R., Reid S., and Greenfield P.F. (1992) Improved production of recombinant proteins by the baculovirus expression system using nutrient enriched serum free media. In: Baculovirus and Recombinant Protein Production Processes. Eds. J.M. Vlak, E.J. Schlaeger and A.R. Bernard, Editiones Roche, Basel, Switzerland, 297–303.
Reed L.J., Muench H. (1938) A simple method of estimating fifty percent endpoints. American Journal of Hygiene, 27: 493–497.
Author information
Authors and Affiliations
Editor information
Rights and permissions
Copyright information
© 1994 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Radford, K.M., Reid, S., Greenfield, P.F. (1994). Enhanced Productivity in Insect Cell Culture by Control of the Chemical Environment. In: Kobayashi, T., Kitagawa, Y., Okumura, K. (eds) Animal Cell Technology: Basic & Applied Aspects. The Sixth International Meeting of Japanese Association for Animal Cell Technology JAACT’93, vol 6. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0848-5_29
Download citation
DOI: https://doi.org/10.1007/978-94-011-0848-5_29
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-4366-3
Online ISBN: 978-94-011-0848-5
eBook Packages: Springer Book Archive