Optimization of rAd5 vectored Newcastle vaccine production in HEK293 at high cell densities


Newcastle disease (ND) is a highly contagious and often severe global spread syndrome that affects birds including domestic poultry. It is caused by a virus belonging to the paramyxoviridae family. Indeed, an outbreak of ND can be quite severe, and is considered as a constant threat to the industry and food security worldwide. The disease can be controlled through the administration of effective vaccines. Immunizations with inactivated or live vaccines, although protective, have some eminent disadvantages. The aims of this work is the development of a vectored vaccine using a non-replicative human adenovirus vector, expressing the F antigen from Newcastle Disease Virus (rAd-F-ND) in bioreactor. The recombinant vaccine is produced using the HEK293 cell line. HEK293 cultures were carried out in suspension, first in shake flasks and then in stirred bioreactor at 37°C, 5% CO2 and 150 rpm in chemically defined media. The virus titers were determined by qPCR. To improve rAd-F-ND virus productions in HEK-293 cells, we studied the effects of the following parameters in shake flask cultures: culture media (Hycell Trans FX-H and Xell-GM), cell density, multiplicity of infection (MOI) and feed (Xell-FS , Xell-GM and cell Boost 5).

Vaccine Technology VIII, ECI Symposium Series 2022