POSTER

Suspension-based adenovirus production process development and scale-up in scale-X™ fixed-bed bioreactors

Cedric Meeus, Thomas Bouckenooghe, Emilie Gateau, Benedicte Dehouck, Sylvain Fauquenoy, Marco Soriani, Maria Ferraiuolo, Alessia Noto, Stefano Colloca, Hanna P. Lesch

Abstract

Suspension-cultured HEK293 cells and serum-free conditions have become an industrial standard for adenovirus production. To maximize the virus titers, a good cell state and high cell density are required. The scale-X™ fixed-bed bioreactor enables high cell density cultivation of surface-dependent and suspension cell lines. In this project, the manufacturing of a non-replicative Gorilla Adenovirus vector (GRAd, proprietary of ReiThera) in suspension cells was transferred to the scale-X™ architecture.

After process transfer, linear scale-up and optimization, productivity was increased reaching in scale-X™ nitro 200m² large scale bioreactor 9.0 x 10⁴ VP/cell for a total yield of 1.75 x 10¹⁷ VPs.

For the first time adenovirus was produced using suspension cells in NevoLine™ upstream platform with integrated scale-X™ nitro bioreactor with results proving straightforward process transfer and scalability.

  • Successful adaptation of suspension-cultured HEK293 cells in fixed-bed architecture;
  • Process transfer, optimization and linear scale-up to manufacturing scale within 9 months;
  • Consistency in metabolites profiles and increase of 3.6-fold of the final cell-specific titer;
  • Reduced working volume compared to suspension systems.