High Cell Density Perfusion Process of Quail Cells Producing Oncolytic rVSV-NDV

Area of Science:

• Oncolytic virotherapy
• Bioprocess engineering
• Cancer treatment

Background:

• Oncolytic viruses show promise for cancer treatment, with rVSV-NDV demonstrating efficacy.
• Efficient production of infectious viruses is essential for virotherapy, but current methods face challenges.
• Good Manufacturing Practice (GMP)-compliant CCX.E10 cells are a potential high-titer producer for rVSV-NDV.

Purpose of the Study:

• To optimize the production of rVSV-NDV using CCX.E10 cells in a perfusion culture system.
• To evaluate the efficacy of semi-perfusion and full perfusion processes for high-density cell culture and virus production.
• To assess the performance of tangential flow depth filtration (TFDF) for cell retention and continuous virus harvesting.

Main Methods:

• Testing quail-originated CCX.E10 cells in semi-perfusion culture with various media and cell densities.
• Implementing a full perfusion process in a 3 L bioreactor utilizing a TFDF device for cell retention.
• Utilizing an integrated depth filter (2-5 µm pore size) for efficient cell retention and continuous virus harvesting.

Main Results:

• The optimized perfusion process achieved viable cell concentrations (VCCs) up to 20.6 × 10⁶ cells/mL with 99.9% cell retention.
• Production of 1.33 × 10⁹ TCID₅₀/mL infectious rVSV-NDV was achieved.
• A 5-fold increase in space-time yield (STY) was observed compared to the control batch process.

Conclusions:

• Perfusion culture significantly enhances rVSV-NDV production yields compared to traditional batch methods.
• GMP-compliant CCX.E10 cells combined with perfusion bioreactors offer a scalable solution for oncolytic virus manufacturing.
• This optimized bioprocess addresses critical production bottlenecks, facilitating clinical translation and broader accessibility of oncolytic virus immunotherapy.