A high-yielding serum-free, suspension cell culture process to manufacture recombinant adenoviral vectors for gene therapy
We have developed an efficient, reproducible, and scaleable cell culture process for a recombinant adenoviral vector expressing therapeutic transgenes for clinical trials. HEK 293 cells – which support the propagation of E1 deficient adenovirus – were first adapted to serum free media and suspension growth. Subsequent studies focused on the infection, virus production and harvest from suspension culture bioreactors. Future studies are planned to address the kinetics of adenovirus production in HEK 293 as well as in other cell lines.
Unable to display preview. Download preview PDF.
- Crystal, RG (1995) Transfer of genes to humans: early lessons and obstacles to success. Science 270: 404-410.Google Scholar
- Graham FL and Prevec L (1991) Manipulation of adenovirus vectors. In: Murray EJ (ed.) Methods in Molecular Biology. Vol. 7, The Humana Press, Clifton, NJ, pp. 109-128.Google Scholar
- Hehir K, Keegan J, Martin J, Pratt D, Auger C, Davis M, Everton M, Narayana R and Karey K (1998) Production issues for adenoviral gene therapy vectors. Presented at Cell Culture Engineering VI, February 7-12, 1998, San Diego, CA.Google Scholar
- Marcel T and Grausz JD (1997) The TMC worldwide gene therapy enrollment report, end 1996. Human Gene Therapy 8: 775-800.Google Scholar
- Nielsen LK, Smyth GK and Greenfield PF (1992) Accuracy of the endpoint assay for virus titration. Cytotechnology 8: 231-236.Google Scholar
- Roth G, Smith C, Schoofs GM, Montgomery TJ, Ayala JL and Horwitz JI (1997) Using an external vortex flow filtration device for perfusion cell culture. BioPharm 10: 30-35.Google Scholar
- Shabram PW, Giroux DD, Goudreau AM, Gregory RJ, Horn MT, Huyghe BG, Liu X, Nunnally MH, Sugarman BJ and Sutjipto S (1997) Analytical anion-exchange HPLC of recombinant type-5 adenoviral particles. Human Gene Therapy 8: 453-465.Google Scholar