Abstract
Recombinant adenoviruses (AdV) are highly efficient at gene transfer for a broad spectrum of cell types and species. They became one of the vectors of choice for gene delivery and expression of foreign proteins in gene therapy and vaccination purposes. To meet the need of significant amounts of adenoviral vectors for preclinical and possibly clinical uses, scalable and reproducible production processes are required.
In this chapter, we review processes used for scalable production of two types of first generation (E1-deleted) adenoviral vectors (Human and Canine) using stirred tank bioreactors. The production of adenovirus vectors using either suspension (HEK 293) or anchorage-dependent cells (MDCK-E1) are described to exemplify scalable production processes with different cell-culture types. The downstream processes will be covered in the next chapter.
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References
McConnell MJ, Imperiale MJ (2004) Biology of adenovirus and its use as a vector for gene therapy. Hum Gene Ther 15(11):1022–1033
Tatsis N, Ertl HC (2004) Adenoviruses as vaccine vectors. Mol Ther 10(4):616–629
Dormond E, Perrier M, Kamen A (2009) From the first to the third generation adenoviral vector: what parameters are governing the production yield? Biotechnol Adv 27(2):133–144
Sheets RL, Stein J, Bailer RT, Koup RA, Andrews C, Nason M, He B, Koo E, Trotter H, Duffy C, Manetz TS, Gomez P (2008) Biodistribution and toxicological safety of adenovirus type 5 and type 35 vectored vaccines against human immunodeficiency virus-1 (HIV-1), Ebola, or Marburg are similar despite differing adenovirus serotype vector, manufacturer’s construct, or gene inserts. J Immunotoxicol 5(3):315–335
Shott JP, McGrath SM, Pau MG, Custers JH, Ophorst O, Demoitie MA, Dubois MC, Komisar J, Cobb M, Kester KE, Dubois P, Cohen J, Goudsmit J, Heppner DG, Stewart VA (2008) Adenovirus 5 and 35 vectors expressing Plasmodium falciparum circumsporozoite surface protein elicit potent antigen-specific cellular IFN-gamma and antibody responses in mice. Vaccine 26(23):2818–2823
Kremer EJ, Boutin S, Chillon M, Danos O (2000) Canine adenovirus vectors: an alternative for adenovirus-mediated gene transfer. J Virol 74(1):505–512
Perreau M, Kremer EJ (2005) Frequency, proliferation, and activation of human memory T cells induced by a nonhuman adenovirus. J Virol 79(23):14595–14605
Soudais C, Boutin S, Kremer EJ (2001) Characterization of cis-acting sequences involved in canine adenovirus packaging. Mol Ther 3(4):631–640
Bru T, Salinas S, Kremer EJ (2010) An update on canine adenovirus type 2 and its vectors. Viruses 2(9):2134–2153
Fallaux FJ, Bout A, van der Velde I, van den Wollenberg DJ, Hehir KM, Keegan J, Auger C, Cramer SJ, van Ormondt H, van der Eb AJ, Valerio D, Hoeben RC (1998) New helper cells and matched early region 1-deleted adenovirus vectors prevent generation of replication-competent adenoviruses. Hum Gene Ther 9(13):1909–1917
Schiedner G, Hertel S, Kochanek S (2000) Efficient transformation of primary human amniocytes by E1 functions of Ad5: generation of new cell lines for adenoviral vector production. Hum Gene Ther 11(15):2105–2116
Cote J, Garnier A, Massie B, Kamen A (1998) Serum-free production of recombinant proteins and adenoviral vectors by 293SF-3F6 cells. Biotechnol Bioeng 59(5):567–575
Nadeau I, Gilbert PA, Jacob D, Perrier M, Kamen A (2002) Low-protein medium affects the 293SF central metabolism during growth and infection with adenovirus. Biotechnol Bioeng 77(1):91–104
Ferreira TB, Ferreira AL, Carrondo MJ, Alves PM (2005) Two different serum-free media and osmolality effect upon human 293 cell growth and adenovirus production. Biotechnol Lett 27(22):1809–1813
Maranga L, Aunins JG, Zhou W (2005) Characterization of changes in PER.C6 cellular metabolism during growth and propagation of a replication-deficient adenovirus vector. Biotechnol Bioeng 90(5):645–655
Mendonca RZ, Prado JCM, Pereira CA (1999) Attachment, spreading and growth of VERO cells on microcarriers for the optimization of large scale cultures. Bioprocess Biosyst Eng 20:565–571
Wu SC, Huang GY, Liu JH (2002) Production of retrovirus and adenovirus vectors for gene therapy: a comparative study using microcarrier and stationary cell culture. Biotechnol Prog 18(3):617–622
Kamen A, Henry O (2004) Development and optimization of an adenovirus production process. J Gene Med 6(Suppl 1):S184–S192
Silva AC, Peixoto C, Lucas T, Kuppers C, Cruz PE, Alves PM, Kochanek S (2010) Adenovirus vector production and purification. Curr Gene Ther 10(6):437–455
Fernandes P, Peixoto C, Santiago VM, Kremer EJ, Coroadinha AS, Alves PM (2013) Bioprocess development for canine adenovirus type 2 vectors. Gene Ther 20:353–360
Sandhu KS, Al-Rubeai M (2008) Monitoring of the adenovirus production process by flow cytometry. Biotechnol Prog 24(1):250–261
Segura MM, Puig M, Monfar M, Chillon M (2012) Chromatography purification of canine adenoviral vectors. Hum Gene Ther Methods 23:182–197
Peixoto C, Ferreira TB, Sousa MF, Carrondo MJ, Alves PM (2008) Towards purification of adenoviral vectors based on membrane technology. Biotechnol Prog 24(6):1290–1296
Croyle MA, Roessler BJ, Davidson BL, Hilfinger JM, Amidon GL (1998) Factors that influence stability of recombinant adenoviral preparations for human gene therapy. Pharm Dev Technol 3(3):373–383
Obenauer-Kutner LJ, Ihnat PM, Yang TY, Dovey-Hartman BJ, Balu A, Cullen C, Bordens RW, Grace MJ (2002) The use of field emission scanning electron microscopy to assess recombinant adenovirus stability. Hum Gene Ther 13(14):1687–1696
Croyle MA, Cheng X, Wilson JM (2001) Development of formulations that enhance physical stability of viral vectors for gene therapy. Gene Ther 8(17):1281–1290
Evans RK, Nawrocki DK, Isopi LA, Williams DM, Casimiro DR, Chin S, Chen M, Zhu DM, Shiver JW, Volkin DB (2004) Development of stable liquid formulations for adenovirus-based vaccines. J Pharm Sci 93(10):2458–2475
Cruz PE, Silva AC, Roldao A, Carmo M, Carrondo MJ, Alves PM (2006) Screening of novel excipients for improving the stability of retroviral and adenoviral vectors. Biotechnol Prog 22(2):568–576
Rexroad J, Wiethoff CM, Green AP, Kierstead TD, Scott MO, Middaugh CR (2003) Structural stability of adenovirus type 5. J Pharm Sci 92(3):665–678
Altaras NE, Aunins JG, Evans RK, Kamen A, Konz JO, Wolf JJ (2005) Production and formulation of adenovirus vectors. Adv Biochem Eng Biotechnol 99:193–260
Rexroad J, Evans RK, Middaugh CR (2006) Effect of pH and ionic strength on the physical stability of adenovirus type 5. J Pharm Sci 95(2):237–247
Rexroad J, Martin TT, McNeilly D, Godwin S, Middaugh CR (2006) Thermal stability of adenovirus type 2 as a function of pH. J Pharm Sci 95(7):1469–1479
Cruz PE, Cunha A, Peixoto CC, Clemente J, Moreira JL, Carrondo MJ (1998) Optimization of the production of virus-like particles in insect cells. Biotechnol Bioeng 60(4):408–418
Maranga L, Cunha A, Clemente J, Cruz P, Carrondo MJ (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(1):55–64
Acknowledgments
The authors acknowledge the financial support from Fundação para a Ciência e Tecnologia (FCT), Portugal (projects PTDC/EBB-BIO/119501/2010 and PTDC/EBB-BIO/118615/2010) and the FP7 EU project BrainCAV (HEALTH-HS_2008_222992). A.C. Silva and P. Fernandes acknowledge the FCT for the Ph.D. grants SFRH/BD/45786/2008 and SFRH/BD/70810/2010, respectively.
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Silva, A.C., Fernandes, P., Sousa, M.F.Q., Alves, P.M. (2014). Scalable Production of Adenovirus Vectors. In: Chillón, M., Bosch, A. (eds) Adenovirus. Methods in Molecular Biology, vol 1089. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-679-5_13
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DOI: https://doi.org/10.1007/978-1-62703-679-5_13
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