Abstract
Many types of disposable bioreactors for protein expression in insect and mammalian cells are now available. They differ in design, capacity, and sensor options, with many selections available for either rocking platform, orbitally shaken, pneumatically mixed, or stirred-tank bioreactors lined with an integral disposable bag (Shukla and Gottschalk, Trends Biotechnol 31(3):147–154, 2013). WAVE Bioreactors™ were among the first disposable systems to be developed (Singh, Cytotechnology 30:149–158, 1999). Since their commercialization in 1999, Wave Bioreactors have become routinely used in many laboratories due to their ease of operation, limited utility requirements, and protein expression levels comparability to traditional stirred-tank bioreactors. Wave Bioreactors are designed to use a presterilized Cellbag™, which is attached to a rocking platform and inflated with filtered air provided by the bioreactor unit. The Cellbag can be filled with medium and cells and maintained at a set temperature. The rocking motion, which is adjusted through angle and rock speed settings, provides mixing of oxygen (and CO2, which is used to control pH in mammalian cell cultures) from the headspace created in the inflated Cellbag with the cell culture medium and cells. This rocking motion can be adjusted to prevent cell shear damage. Dissolved oxygen and pH can be monitored during scale-up, and samples can be easily removed to monitor other parameters. Insect and mammalian cells grow very well in Wave Bioreactors (Shukla and Gottschalk, Trends Biotechnol 31(3):147–154, 2013). Combining Wave Bioreactor cell growth capabilities with recombinant baculoviruses engineered for insect or mammalian cell expression has proven to be a powerful tool for rapid production of a wide range of proteins.
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
Shukla AA, Gottschalk U (2013) Single-use disposable technologies for biopharmaceutical manufacturing. Trends Biotechnol 31(3):147–154
Singh V (1999) Disposable bioreactor for cell culture using wave-induced agitation. Cytotechnology 30:149–158
Eibl R, Kaiser S, Lobriser R et al (2010) Disposable bioreactors: the current state-of-the-art and recommended applications in biotechnology. Appl Microbiol Biotechnol 86:41–49
Weber W, Weber E, Geisse S et al (2002) Optimization of protein expression and establishment of the Wave Bioreactor for Baculovirus/insect cell culture. Cytotechnology 38:77–85
Kadwell SH, Hardwicke PI (2007) Production of baculovirus-expressed recombinant proteins in wave bioreactors. In: Murhammer D (ed) Baculovirus and insect cell expression protocols, 2nd edn. Humana Press, Totowa, NJ, pp 247–266
Wang L, Hu H, Yang J, Wang F, Kaisermayer C, Zhou P (2012) High yield of human monoclonal antibody produced by stably transfected Drosophila Schneider 2 cells in perfusion culture using wave bioreactor. Mol Biotechnol 52(2):170–179
Hami LS, Green C, Leshinsky N et al (2004) GMP production and testing of Xcellerated T cells for the treatment of patients with CLL. Cytotherapy 6(6):554–562
Somerville RP, Devillier L, Parkhurst MR et al (2012) Clinical scale rapid expansion of lymphocytes for adoptive cell transfer therapy in the WAVE® bioreactor. J Transl Med 10:69
van der Loo JC, Swaney WP, Grassman E et al (2012) Scale-up and manufacturing of clinical-grade self-inactivating γ-retroviral vectors by transient transfection. Gene Ther 19(3):246–254
Kwon JY, Yang YS, Cheon SH et al (2013) Bioreactor engineering using disposable technology for enhanced production of hCTLA4Ig in transgenic rice cell cultures. Biotechnol Bioeng 110:2412–2424. doi:10.1002/bit.24916
Dalton JP, Demanga CG, Reiling SJ et al (2012) Large-scale growth of the Plasmodium falciparum malaria parasite in a wave bioreactor. Int J Parasitol 42(3):215–220
Mahajan E, Matthews T, Hamilton R et al (2010) Use of disposable reactors to generate inoculum cultures for E. coli production fermentations. Biotechnol Prog 26(4):1200–1203
Rao G, Moreira A, Brorson K (2009) Disposable bioprocessing: the future has arrived. Biotechnol Bioeng 102(2):348–560
Condreay JP, Witherspoon SM, Clay WC et al (1999) Transient and stable gene expression in mammalian cells transduced with a recombinant baculovirus vector. Proc Natl Acad Sci U S A 96:127–132
Ames RS, Kost TA, Condreay JP (2007) BacMam technology and its application to drug discovery. Expert Opin Drug Discov 2(12):1669–1681
Ramos L, Kopec LA, Sweitzer SM et al (2002) Rapid expression of recombinant proteins in modified CHO cells using the baculovirus system. Cytotechnology 38(1-3):37–41
Hu YC, Tsai CT, Chang YJ et al (2003) Enhancement and prolongation of baculovirus-mediated expression in mammalian cells: focuses on strategic infection and feeding. Biotechnol Prog 19(2):373–379
Wulhfard S, Tissot S, Bouchet S et al (2008) Mild hypothermia improves transient gene expression yields several fold in Chinese hamster ovary cells. Biotechnol Prog 24(2):458–465
Scott MJ, Modha SS, Rhodes AD et al (2007) Efficient expression of secreted proteases via recombinant BacMam virus. Protein Expr Purif 52:104–116
Condreay JP, Ames RS, Hassan NJ et al (2006) Baculoviruses and mammalian cell-based assays for drug screening. Adv Virus Res 68:255–286
Chiocca S, Baker A, Cotten M (1997) Identification of a novel antiapoptotic protein, GAM-1, encoded by the CELO adenovirus. J Virol 71(4):3168–3177
Chiocca S, Kurtev V, Colombo R et al (2002) Histone deacetylase 1 inactivation by an adenovirus early gene product. Curr Biol 12(7):594–598
Hacker DL, Derow E, Wurm FM (2005) The CELO adenovirus Gam1 protein enhances transient and stable recombinant protein expression in Chinese hamster ovary cells. J Biotechnol 117(1):21–29
Gallimore PH, Turnell AS (2001) Adenovirus E1A: remodelling the host cell, a life or death experience. Oncogene 20(54):7824–7835
Cockett MI, Bebbington CR, Yarranton GT (1991) The use of engineered E1A genes to transactivate the hCMV-MIE promoter in permanent CHO cell lines. Nucleic Acids Res 19(2):319–325
Kemp CW, Gugel A, Birch A (2012) Transient expression of recombinant immunoglobulin in HEK-293 and CHO-S cells using BacMam transduction. BioProcess J 11(2):4–12
Janakiraman V, Forrest WF, Seshagiri S (2006) Estimation of baculovirus titer based on viable cell size. Nat Protoc 1(5):2271–2276
Acknowledgements
Condreay J.P., Clay W.C., and Kost T.A. for the details on the CHO-GE cell line construction.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Kadwell, S.H., Overton, L.K. (2016). Protein Expression in Insect and Mammalian Cells Using Baculoviruses in Wave Bioreactors. In: Murhammer, D. (eds) Baculovirus and Insect Cell Expression Protocols. Methods in Molecular Biology, vol 1350. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3043-2_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3043-2_12
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3042-5
Online ISBN: 978-1-4939-3043-2
eBook Packages: Springer Protocols