Process intensification for lentiviral vector manufacturing using tangential flow depth filtration.

Area of Science:

• Biotechnology
• Process Engineering
• Gene Therapy Manufacturing

Background:

• Gene therapies require scalable and cost-effective manufacturing of viral vectors.
• Process intensification is a key strategy to increase viral vector yield per batch.
• Lentiviral vectors are crucial for CAR T and TCR cell therapies.

Purpose of the Study:

• To investigate process intensification for lentiviral vector manufacturing.
• To evaluate tangential flow depth filtration for enabling perfusion and cell expansion.
• To determine the potential dose output of an intensified lentiviral vector production process.

Main Methods:

• Implemented perfusion in a production bioreactor using a stable producer cell line.
• Utilized hollow-fiber depth filters (polypropylene, 2-4 μm channels) for tangential flow depth filtration.
• Operated the intensified process at a 200-L scale in suspension culture.

Main Results:

• Tangential flow depth filtration enabled perfusion, leading to expanded cell densities.
• The depth filters demonstrated high capacity, extended functional life, and efficient separation of lentiviral vectors.
• The intensified process is anticipated to yield approximately 10,000 doses per batch.

Conclusions:

• Process intensification with tangential flow depth filtration is a viable strategy for increasing lentiviral vector production.
• This approach can significantly enhance the accessibility and affordability of gene therapies.
• The technology shows promise for meeting the large-scale demand for CAR T and TCR therapies.