Baculovirus-insect cell expression system

Andersen, J.N., "Temperature Effect on Recombinant Protein Production Using a Baculovirus/Insect Cell Expression System", Diploma Thesis, University of Calgary and Technical University of Denmark, 1995. 

Jarvis, D.L. (2003) Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology, 310 (1), 1-7. 

Caron AW, Archambault J, Massie B (1990), High-level recombinant protein production in bioreactors using the baculovirus-insect cell expression system, Biotechnol. Bioeng. 36 1133-1140. 

Betenbaugh M J, Ailor E, Whiteley E, Hinderliter P, Hsu T A (1996). Chaperone and foldase coexpression in the baculovirus-insect cell expression system. Cytotechnology 20 149-159. 

Mao W, Berenbaum MR, Schuler MA (2008) Modifications in the N-terminus of an insect P450 enhance production of catalytically active protein in baculovirus-Sf9 cell expression systems. Insect Biochem Mol Biol 38 66 75... 

Insect cell expression Baculovirus High-level expression Formation of disulphides Glycosylation Glycosylated product may differ from mammalian systems Not necessarily fully functional... 

Weber, W, Weber, E., Geisse, S. and Memmert, K. (2002). Optimisation of protein expression and establishment of the Wave Bioreactor system for Baculovirus/insect cell culture. Cytotechnology 38, 77-85. 

In summary, we were able to demonstrate the expression of large amounts of a functional sGC using the baculovirus-insect cell system. Supplementation of expressed sGC with heme or the heme analog zinc protoporphyrin IX drastically stimulates or inhibits NO-mediated enzymatic activity. This high expression level of sGC should facilitate future physicochemical and structural studies. 

Recombinant baculoviruses and their lepidopteran insect cell hosts have become one of the most widely used expression systems for the production of recombinant proteins [49, 50]. Manipulation of the system is straightforward, and high yields of recombinant material can be readily obtained in a matter of a few weeks compared with the several months that are required with stable animal cell expression systems. Generally, the insect-produced proteins are intended for research applications rather than as pharmaceuticals. The glycans on insect proteins differ significantly from those found on animal cell proteins, and this has limited the development of an attractive expression system for production of human therapeutics. 

Funk CD, Gunne H, Steiner H, Izumi T, Samuelsson B. Native and mutant 5-lipoxygenase expression in a baculovirus insect cell system. Proc Natl Acad Sci USA 1989 86 2592-2596. 

Insect cell systems represent multiple advantages compared with mammalian cell cultures (1) they are easier to handle (Table 2.1) (2) cultivation media are usually cheaper (3) they need only minimum safety precautions, as baculovirus is harmless for humans (4) they provide most higher eukaryotic posttranslational modifications and heterologous eukaryotic proteins are usually obtained in their native conformation (5) the baculovirus system is easily scalable to the bioreactor scale. However, because of the viral nature of the system, continuous fermentation for transient expression is not possible - the cells finally die. 

Zhang, F., Saarinen, M.A., Itle, L.J. et al. (2002) The effect of dissolved oxygen (DO) concentration on the glycosylation of recombinant protein produced by the insect cell-baculovirus expression system. Biotechnology and Bioengineering, 11 (2), 219-224. 

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