Monoclonal Antibody (mAb) Production from Continuous Hybridoma Culture for 100 Days in the LAMBDA MINIFOR Bioreactor
Continuous bioprocessing is increasingly adopted to improve:
- Productivity
- Product consistency
- Operational efficiency
However, it requires a system capable of maintaining stable conditions for sensitive mammalian cells over long periods.
A study conducted at the Center for Genetic Engineering and Biotechnology (CIGB), demonstrated that the LAMBDA MINIFOR bioreactor provides the required stability and control for long-term mammalian cell cultivation.
The system successfully supported a continuous hybridoma culture for 100 days in protein-free medium – without performance drift or contamination
Reference: González M., Aragón H., Medel M., Hernández D., Ferro W., et al. (2024). Continuous culture of mouse hybridoma cells for 100 Days in the LAMBDA MINIFOR bioreactor. Biotecnología Aplicada, 41(2): 2211–2218.
Study Overview
CB.Hep-1 hybridoma cells were cultivated continuously in the MINIFOR bioreactor under controlled conditions:
- Temperature: 36–37 °C
- pH: 6.8–7.2
- Low-shear conditions using biomimetic fishtail agitation
The gentle mixing design minimizes shear stress—an important factor for maintaining hybridoma viability and productivity over long durations.
Aeration was maintained at low levels without requiring CO₂ supplementation.
Daily medium renewal was performed, enabling stable long-term operation.
Hybridoma cultures are particularly sensitive to long-term instability, including genetic drift and productivity loss. Therefore, maintaining stable performance over extended periods is a critical benchmark for bioreactor reliability.
Continuous Operation
Continuous operation began on Day 6 and was maintained for 100 days with up to 19% daily volume replacement, ensuring steady-state conditions.
This enabled stable steady-state conditions throughout the cultivation period.
Performance Results
The LAMBDA MINIFOR maintained stable cell culture performance throughout the entire experiment:
- Average viability: 82.5%
- Viable cell density: 1.1 × 10⁶ cells/mL
Monoclonal antibody production:
- Mean concentration: 122.7 mg/L
- Specific productivity: 25.7 µg/10⁶ cells/day
Overall, the culture produced 12.5 g of monoclonal antibody, with 10.1 g recovered as highly purified product.
Product Quality and Stability
Product quality remained highly consistent throughout the entire process:
- Purity > 99%
- No aggregation
- Stable antigen-binding specificity
Importantly, extended continuous cultivation did not negatively affect molecular integrity or biological activity.
The monoclonal antibody (mAb):
- retained structural integrity
- maintained affinity
- showed no degradation
This demonstrates the suitability of the MINIFOR for applications requiring long-term product consistency, including hybridoma-based production and continuous perfusion processes.
Scale-Up Potential
Based on MINIFOR data, large-scale performance was modeled:
- 600 L (no cell retention) – Continuous – 2,025 g mAb/year
- 50 L (with cell retention) – Continuous – 1,378 g mAb/year
These projections highlight the role of the MINIFOR as a reliable scale-down model for process development and optimization prior to industrial implementation.
Conclusion
The 100-day CB.Hep-1 hybridoma study demonstrates that the LAMBDA MINIFOR bioreactor provides:
- Stable long-term operation
- Precise environmental control
- Gentle mixing for sensitive cells
- Consistent product quality
The system is particularly suited for applications that can maintain stable biological conditions over extended periods.
The MINIFOR provides a robust and reliable platform for mammalian cell culture, monoclonal antibody (mAb) production, and continuous process development.
Interested in applying continuous culture in your lab?
Contact us to discuss your application, configuration options, or to arrange a technical discussion:
Email us: sales@lambda-instruments.com