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Published on: July 11, 2013
Process Intensification for Recombinant Marburg Virus Glycoprotein Production Using Drosophila S2 Cells
Sven Göbel1, Ludwig Mayerlen2, Isabelle Yazel Eiser2
1Bioprocess Engineering Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr Magdeburg Germany.
Abstract:
Marburg marburgvirus (MARV) is a highly virulent human pathogen with limited therapeutic options. Recombinant MARV glycoprotein (GP) produced in Drosophila Schneider 2 (S2) cells has been extensively investigated as potential vaccine antigen with promising efficacy demonstrated in nonhuman primate models. However, the existing production process for MARV-GP involving static batch cell cultures with limited scalability and process control show lower than desirable yields. Here, we assessed various process intensification strategies in single-use orbital shaken bioreactors (OSBs) or rocking bioreactors (WAVE) and report maximum viable cell concentrations (VCCs) of 31.6 × 106 cells/mL in batch, 69.5 × 106 cells/mL in fed-batch (FB), and up to 210.0 × 106 cells/mL in perfusion mode. By changing from a glucose-only feed to a CellBoost5 feed, MARV-GP yields were increased by over two-fold. Implementation of perfusion cultures achieved a peak MARV-GP concentration of 57.4 mg/L and a 540% higher space-time yield compared to the FB process in the 50 L WAVE system. However, maximum cell-specific productivities were achieved at a VCC of 85 × 106 cells/mL and decreased with increasing cell concentrations. Glycoanalysis revealed a uniform paucimannosidic N-glycan profile, predominantly α-1,6-core-fucosylated Man3F (F(6)M3) structures, across all production modes. Notably, transitioning pH control from CO2 to phosphoric acid shifted glycan profiles toward higher mannose forms, highlighting the influence of culture conditions on glycosylation.
Insights
Process intensification strategies significantly improved Marburg marburgvirus glycoprotein (MARV-GP) production yields. Perfusion cultures in bioreactors achieved higher concentrations and space-time yields compared to fed-batch, optimizing vaccine antigen manufacturing.
Area of Science:
- Biotechnology and Bioprocessing
- Vaccine Antigen Production
- Virology
Background:
- Marburg marburgvirus (MARV) is a dangerous pathogen with few treatment options.
- Recombinant MARV glycoprotein (GP) from Drosophila Schneider 2 (S2) cells shows vaccine potential but current production yields are suboptimal.
- Existing static batch cultures limit scalability and process control for MARV-GP.
Purpose of the Study:
- To assess process intensification strategies for enhanced MARV-GP production.
- To evaluate scalability and yield improvements using single-use bioreactors.
- To analyze the impact of culture conditions on MARV-GP glycosylation.
Main Methods:
- MARV-GP production was evaluated in single-use orbital shaken bioreactors (OSBs) and rocking bioreactors (WAVE).
- Process intensification strategies included fed-batch (FB) and perfusion culture modes.
- Different feed formulations (glucose vs. CellBoost5) and pH control methods (CO2 vs. phosphoric acid) were tested.
Main Results:
- Maximum viable cell concentrations (VCCs) reached 210.0 × 10^6 cells/mL in perfusion mode.
- Perfusion cultures yielded a peak MARV-GP concentration of 57.4 mg/L, a 540% increase in space-time yield over FB.
- CellBoost5 feed doubled MARV-GP yields; optimal cell-specific productivity occurred at 85 × 10^6 cells/mL.
Conclusions:
- Process intensification, particularly perfusion culture, significantly enhances MARV-GP production in bioreactors.
- Culture conditions, including feed type and pH control, influence MARV-GP yield and glycosylation profiles.
- Optimized bioreactor processes offer a scalable and efficient method for producing MARV-GP vaccine antigens.
