Perfusion process with tangential flow filtration for oncolytic VSV-GP production

Area of Science:

• Biotechnology
• Virology
• Chemical Engineering

Background:

• Oncolytic vesicular stomatitis virus (VSV)-GP shows therapeutic potential for cancer treatment.
• Achieving high virus titers is crucial for clinical efficacy but challenging in manufacturing.
• Perfusion cultivation at high cell densities is a promising strategy to boost production titers.

Purpose of the Study:

• To enhance the production titer of VSV-GP using suspension human embryonic kidney 293 (HEK293) cells.
• To optimize a perfusion cultivation process incorporating tangential flow filtration (TFF) and virus retention.
• To identify and evaluate critical process parameters influencing VSV-GP yield.

Main Methods:

• Utilized I-optimal design of experiments (DoE) to assess six critical process parameters.
• Employed suspension HEK293 cells in a perfusion system with TFF and virus retention.
• Investigated the impact of seeding cell density, perfusion pause duration, exchange rate, and shear stress.

Main Results:

• Increasing seeding cell density improved infectious titer up to 46.6 × 10^6 cells mL^-1.
• Minimizing perfusion pause (1.1-1.3 h) and using a higher exchange rate (0.045-0.051 nL cell^-1 d^-1) were beneficial.
• Optimal crossflow rate (44-55 mL min^-1) minimized shear stress sensitivity.
• Achieved a 17-fold increase in titer (up to 5.1 × 10^10 TCID50 mL^-1) compared to batch cultivation.

Conclusions:

• Optimized perfusion cultivation significantly enhances VSV-GP production titers.
• High cell densities and controlled perfusion parameters are key to maximizing virus yield.
• Perfusion cultivation with virus retention is an effective technology for improving VSV-GP manufacturing.