High yield sterile filtration process for highly concentrated lentiviral vectors

Area of Science:

• Biopharmaceutical Manufacturing
• Gene Therapy
• Filtration Technology

Background:

• Biopharmaceutical production requires stringent quality control, often involving sterile filtration.
• Lentiviral vectors (LVVs) used in ex vivo gene therapy face filtration challenges due to their size relative to standard filter pore sizes (0.2 µm).
• High concentrations of LVVs can lead to filter clogging caused by aggregation or impurity attachment.

Purpose of the Study:

• To identify suitable sterile filters for high-titer lentiviral vector (LVV) production.
• To evaluate the impact of endonuclease treatment on filter performance during LVV purification.
• To establish a scalable sterile filtration process for LVV manufacturing.

Main Methods:

• Screening of various filtration materials to assess compatibility with high-titer LVV stocks (up to 1E+9 transducing units/mL).
• Investigation of the effect of upstream endonuclease treatment on filter performance and LVV recovery.
• Small-scale filtration studies (<15 mL) followed by successful scale-up to 100 mL.

Main Results:

• Three filter types were identified as suitable for sterile filtration of concentrated LVV stocks.
• Filtration processes achieved high virus yields (>80%) with the selected filters.
• The filtration method was successfully scaled up from small-scale experiments to a 100 mL process volume.

Conclusions:

• Specific filters enable effective sterile filtration of high-titer lentiviral vectors, a critical step for gene therapy manufacturing.
• The developed filtration process maintains high LVV yield and quality, addressing a key manufacturing challenge.
• This work supports the development of robust and scalable manufacturing processes for LVV-based gene therapies.