Monoclonal Antibody Production from Continuous Hybridoma Culture for 100 Days in the LAMBDA MINIFOR Bioreactor
The LAMBDA MINIFOR laboratory bioreactor has proven its robustness and precision in a groundbreaking 100-day continuous culture of mouse hybridoma cells for monoclonal antibody (mAb) production. Conducted by researchers at the Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba, this study demonstrated the stability of CB.Hep-1 hybridoma cells in protein-free medium using the MINIFOR system. Over the extended run, the culture maintained 82.5% average cell viability and consistently produced high-purity monoclonal antibodies (>98%) without any biochemical or immunological modifications.
These findings confirm that the LAMBDA MINIFOR bioreactor provides reliable control for long-term mammalian cell cultures and serves as an ideal platform for scaling up continuous bioprocesses to industrial levels.
Background
Continuous bioprocessing represents the next evolution in biopharmaceutical manufacturing. It offers reduced downtime, lower production costs, and enhanced product consistency compared to traditional batch or fed-batch modes. However, the key challenge lies in maintaining stable cell performance over extended periods.
Hybridoma-based mAb production, although historically robust, can be prone to genetic drift and productivity loss under prolonged culture conditions. Therefore, maintaining cell line stability and product quality over extended periods is a critical benchmark for evaluating bioreactor performance.
The CIGB team selected the LAMBDA MINIFOR bioreactor—equipped with a biomimetic fishtail agitation system and precise control of temperature, pH, and aeration—to validate long-term hybridoma culture performance.
Experimental Setup
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Cell line: Mouse hybridoma CB.Hep-1 (producer of anti-HBsAg monoclonal antibody)
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Culture medium: Protein-free (PFHM-II) formulation
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LAMBDA MINIFOR Bioreactor configuration:
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Working volume: 2–6 L
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Operation mode: Continuous (after initial fed-batch phase)
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Temperature: 36–37 °C
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pH: 6.8–7.2
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Agitation: 4 Hz (fishtail motion)
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Aeration: 0.01–0.02 L/min (no CO₂ supply needed)
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Continuous operation began on Day 6 and was maintained for 100 days with up to 19% daily volume replacement, ensuring steady-state conditions.
Key Results
| Parameter | Average Value | Comment |
|---|---|---|
| Viable cell density | 1.1 ± 0.2 × 10⁶ cells/mL | Stable throughout 100 days |
| Viability | 82.5 ± 6.7 % | High and consistent |
| Specific production rate | 25.7 ± 9.8 µg mAb/10⁶ cells·day | Comparable to high-performance systems |
| mAb concentration | 122.7 ± 43.6 mg/L | Steady across all harvests |
| Total mAb yield | 12.5 g harvested, 10.1 g purified | Over 98 % purity |
| mAb purity | > 99 % (SDS-PAGE, SEC-HPLC) | No aggregation or degradation |
| Affinity constant | 0.9 × 10¹⁰ M⁻¹ | Unchanged specificity |
The LAMBDA MINIFOR bioreactor maintained consistent performance over the full 100-day run. No loss of productivity or antibody quality was observed—demonstrating excellent process reproducibility and culture stability.
Scale-Up Potential
Based on MINIFOR data, the researchers modeled large-scale continuous operation:
| Scale | Operation Mode | Estimated Annual Yield |
|---|---|---|
| 600 L (no cell retention) | Continuous | 2,025 g mAb/year |
| 50 L (with cell retention) | Continuous | 1,378 g mAb/year |
Such scalability underscores the MINIFOR’s role as an accurate and economical scale-down model for mammalian cell culture optimization—enabling cost-effective transition to pilot and production bioreactors.
Conclusion
The study provides strong evidence that the LAMBDA MINIFOR bioreactor is a robust and reliable system for long-term mammalian cell culture in continuous mode. It maintained cell stability, high productivity, and consistent antibody quality for 100 days—validating its suitability for extended bioprocess research and scale-up.
By minimizing medium consumption and ensuring precise environmental control, the MINIFOR allows scientists to simulate industrial conditions at bench scale, accelerating process development and ensuring scalability with confidence.
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.