Abstract
The levels of many recombinant proteins produced using the baculovirus expression vector (BEV) system are extraordinarily high (1, 2), in many cases approaching 1000 mg/L, the level of polyhedrin expression. Unfortunately, other proteins that have been expressed in the BEV system are produced at much lower levels, which are often >5 mg/L (Table 1). Certainly, vector construction plays a major role in the level of protein expression, and many of the early reports of lower level expression using BEV can probably be traced to the use of suboptimal vectors that lacked sequences important to high level transcription from the polyhedrin promoter 3). However, more recent data on baculovirus expression levels using improved transfer vectors continue to reveal pro teins that are expressed at levels below 5 mg/L (4–6). The questions that remain concern the reasons for such large differences and whether these reasons are in some way related to the insect cell line that is used.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Luckow, V. A. (1990) Cloning and expression of heterologous genes in insect cells with baculovirus vectors, in Recombinant DNA Technology and Applica tions (Ho, C., Prokop, A., and Bajpai, R., eds.), McGraw-Hill, New York.
Luckow, V. A. and Summers, M. D. (1989) High level expression of nonfused foreign genes with Autographa calijornica nuclear polyhedrosis virus expression vectors. Virology 170, 31–39.
Matsuura, Y., Possee, R. D., and Bishop, D. H. L. (1987) Baculovirus expression vectors: The requirements for high level expression of proteins, including glycoproteins. J. Gen. Virol. 67, 1515–1529.
Moore, M. D., Cannon, M. J., Sewall, A., Finlayson, M., Okimoto, M., and Nemerow, G. R. (1991) Inhibition of Epstein-Barr virus infection in vitro and in vivo by soluble CR2 (CD21) containing two short consensus repeats. J. Viral. 65, 3559–3565.
Ruf, W., Miles, D. J., Rehumtulla, A., and Edgington, T. S. (1993) Mutational analysis of receptor and cofactor function of tissue factor. Methods Enzymol. 222, 209–224.
Greenfield, C, Patel, G., Clark, S., Jones, N., and Waterfield, M. D. (1988) Expression of the human EGF receptor with ligand-stimulatable kinase activity in insect cells using a baculovirus vector. EMBO J. 7, 139–146.
Whang, Y., Silberklang, M., Morgan, A., Munshi, S., Lenny, A. B., Ellis, R. W., and Kieff, E. (1987) Expression of the Epstein-Barr virus gp350/220 gene in rodent and primate cells. J. Virol. 61, 1796–1807.
Steiner, H., Pohl, G., Gunne, H., Hellers, M., Elhammer, A., and Hansson, L. (1988) Human tissue-type plasmogen activator synthesized by using a baculovirus vector in insect cells compared with human plasminogen activator produced in mouse cells. Gene 73, 449–457.
Kuroda, K., Geyer, H., Geyer, R., Doerfler, W., and Klenk, H.-D. (1990) The oligosaccharides of influenza virus hemaglutinin expressed in Insect cells by a baculovirus vector. Virology 174, 418–429.
Davidson, D. J., Fraser, M. J., and Castellino, F. J. (1990) Oligosaccharide processing in the expression of human plasminogen cDNA by lepidopteran insect (Spodopterafrugiperda) cells. Biochemistry 29, 5584–5590.
Wojchowslu, D. M., Lorkin, S H., and Sytkowski, A. J. (1987) Active human erythropoietin expressed in insect cells using a baculovirus vector: a role for N-linked oligosaccharide. Biochim. Biophys. Acta 910, 224–232.
Hink, W. F., Thomsen, D. R., Davidson, D. R., Meyer, A. L., and Castellino, F. J. (1991) Expression of three recombinant proteins using baculovirus vectors in 23 insect cell lines. Biotechnol. Prog. 7, 9–14.
Luckow, V. A. and Summers, M. D. (1988) Signals important for high-level expres sion of foreign genes in Autographa culifornica nuclear polyhedrosis virus expression vectors. Virology 167, 56–71.
Jarvis, D. L. and Summers, M. D (1989) Glycosylation and secretion of human tissue plasminogen activator in recombinant baculovirus-infected insect cells. Mol. Cell.Biol. 9, 214–223.
Vernet, T., Tessier, D. C., Richardson, C., Laliberté, F, Khi, H. E, Bell, A. W., Storer, A. C., and Thomas, D. Y. (1990) Secretion of functional papain precursor from insect cells. J. Biol. Chem. 265, 16661–16666.
Wickham, T. J. (1991) Baculovirus-insect cell interactions in producing heterolo gous proteins: Attachment, infection, and expression in different cell lines. Ph. D. Thesis Dissertation, Cornell University.
King, L. A., Mann, S. G., Lawrie, A. M., and Mulshaw, S. H. (1991) Replication of wild-type and recombinant Autographa culifomicu nuclear polyhedrosis virus in a cell line derived from Mumestru brussicue. Virus Res. 19, 93–104.
Wickham, T. J, Davis, T., Granados, R R., Shuler, M. L, and Wood, H. A (1992) Screening of insect cell lines for the production of recombinant proteins and infec tious virus in the baculovirus expression system. Biotechnol. Prog. 8, 391–396.
-Davis, T. R., Wickham, T. J., McKenna, K. A., Granados, R. R., Shuler, M. L., and Wood, H. A. (1993) Comparative recombinant protein production of eight insect cell lines. In Vitro Cell. Dev. Biol. 29A, 388–390.
Davis, T. R., Munkenbeck Trotter, K., Granados, R. R., and Wood, H. A. (1992). Baculovnus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion. Bio/Technology 10, 1148–1150.
Davis, T. R., Shuler, M. L., Granados, R. R., and Wood, H. A. (1993) Comparison of oligosaccharide processing among various insect cell lines expressing a secreted glycoprotein. In Vitro Cell. Dev. Biol. 29A:ll, 842–846.
Wickham, T. J., and Nemerow, G. R. (1993) Optimization of growth methods and recombinant protein production in BTI Tn-5B l-4 insect cells using the baculovirus expression vector. Biotechnol. Prog. 9, 25–30.
Wu, J., King, G., Daugulis, A. J., Faulkner, P., Bone, D. H., and Goosen, M. R. A. (1989) Engineering aspects of insect cell suspension culture: a review. Appl. Microbial. Biotechnol. 32, 249–255.
Lazar, A., Silberstein, L., Reuveny, S., and Mizrahi, A. (1987) Microcarriers as a culturing system of insect cells and insect viruses. Devel. Biol. Standard 64, 315–323.
Shuler, M.L., Cho, T., Wickham, T. J., Ogonah, O., Kool, M., Hammer, D. A., Granados, R. R. and Wood, H. A. (1990) Bioreactor development for production of viral pesticides or heterologous proteins in insect cell cultures. Ann. NY Acud. Sci. 589, 399–422.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Humana Press Inc, Totowa, NJ
About this protocol
Cite this protocol
Wickham, T.J., Nemerow, G.R., Wood, H.A., Shuler, M.L. (1995). Comparison of Different Cell Lines for the Production of Recombinant Baculovirus Proteins. In: Richardson, C.D. (eds) Baculovirus Expression Protocols. Methods in Molecular Biology, vol 39. Humana Press. https://doi.org/10.1385/0-89603-272-8:385
Download citation
DOI: https://doi.org/10.1385/0-89603-272-8:385
Publisher Name: Humana Press
Print ISBN: 978-0-89603-272-9
Online ISBN: 978-1-59259-526-6
eBook Packages: Springer Protocols