Scalable Fed-Batch Bioreactor Process for Lentiviral Vector Production

In cell and gene therapy, the production of lentiviral vectors (LVVs) is a critical component. These vectors are essential for delivering genetic material into cells, a process fundamental to many therapeutic applications. However, scaling up the production of LVVs to meet clinical and commercial demands has been a significant challenge. This case study explores how an effective, scalable fed-batch bioreactor process helps to address these challenges, advancing LVV production.

Scaling up LVV production

A research team at the National Research Council of Canada (NRC) encountered a major obstacle in their goal to produce LVVs at a scale required for clinical trials and commercial therapies. Traditional LVV production methods relied on adherent cell cultures, which were not easily scalable. The process required extensive plasticware and complex handling, creating a bottleneck in producing the necessary quantities of LVVs.

The estimated annual vector requirement for treating just 5,000 pediatric cancer patients was around 3.5×10¹² transduction units. This meant producing hundreds to thousands of liters of LVVs, a task that seemed nearly insurmountable with existing methods.

Developing a scalable and efficient production process

The research team set out with a clear objective: to develop a scalable, efficient, and cost-effective process for producing high-titer LVVs using a cell suspension line. The solution needed to simplify the scale-up process, reduce production costs, and maintain high product quality.

Scalable fed-batch bioreactor process

Key findings

1. Enhanced titer and productivity: The fed-batch process demonstrated a significant increase in LVV-GFP titers compared to batch production. Specifically, the fed-batch regimens (FB R1 and FB R2) achieved up to a 2.6-fold increase in titer, highlighting the process’s efficiency in boosting viral vector yields.
2. Improved cell density and viability: The study reported higher viable cell concentrations in fed-batch cultures, leading to increased specific productivity. This improvement is attributed to controlled culture conditions in the bioreactor, such as pH and dissolved oxygen levels, which are more challenging to maintain in shake flask cultures.
3. Scalability and reproducibility: The process was successfully scaled up from a 4 L bioreactor to a 50 L bioreactor, maintaining consistent metabolite profiles and productivity levels. This scalability is crucial for industrial applications, ensuring that the process can be adapted to larger production volumes without compromising efficiency or product quality.
4. Cost-effectiveness: By optimizing feed additions and reducing the overall culture duration, the fed-batch process offers a cost-effective solution for LVV production. The study’s approach minimizes the use of expensive reagents and reduces the timeline for manufacturing, thereby lowering production costs.

Conclusion

The development of a scalable fed-batch bioreactor process for high-titer LVV production marks an advancement in the field of biotechnology. This study offers a robust framework for producing LVVs at an industrial scale, helping to meet the growing demands of cell and gene therapies. The enhanced titer, improved cell viability, and cost-effectiveness of the process underscore its potential for widespread adoption in clinical research and commercial settings.

To read more about this case study, check out the full paper on BioProcessing Journal.

We’re thrilled that our 50 L DynaDrive Single-Use Bioreactor (S.U.B.) was used in this important study. Visit our website to learn more about the DynaDrive S.U.B.

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