Investigating Ultrafiltration Membranes and Operation Modes for Improved Lentiviral Vector Processing

Area of Science:

• Biotechnology
• Gene Therapy
• Bioprocessing

Background:

• Lentiviral vectors (LVs) are crucial for ex vivo gene therapies.
• LV production faces challenges due to vector instability and fragility.
• Downstream processing optimization is needed to improve LV yield and quality.

Purpose of the Study:

• To investigate the impact of membrane type and filtration device on lentiviral vector ultrafiltration.
• To identify optimal conditions for concentrating infectious LVs and removing impurities.
• To enhance the efficiency and product quality of LV downstream processing.

Main Methods:

• Evaluated nine membrane materials (PES, regenerated cellulose, Hydrosart) with varying molecular weight cutoffs.
• Tested membranes using stirred cells, centrifugal ultrafilters, and crossflow cassettes.
• Assessed performance based on infectious LV particle retention and impurity removal (DNA).

Main Results:

• Reinforced 100 kDa PES and 300 kDa Hydrosart membranes showed superior performance in concentrating infectious LVs and removing DNA.
• Stirred cell operation yielded better results compared to other filtration devices.
• Non-optimized crossflow cassette processes resulted in lower infectious LV recovery.

Conclusions:

• Membrane material and filtration device significantly influence the efficiency of lentiviral vector ultrafiltration.
• Specific membrane types and stirred cell configurations are recommended for improved LV concentration and purity.
• Further optimization of crossflow systems may be necessary to enhance LV recovery.