Process Intensification for Recombinant Marburg Virus Glycoprotein Production Using Drosophila S2 Cells.

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.