Developing an oncolytic Newcastle disease virus production process with EB66 cells

Area of Science:

• Biotechnology
• Virology
• Cancer Research

Background:

• Newcastle disease virus (NDV) is researched as an oncolytic virus for cancer therapy.
• Large-scale manufacturing of high-dose NDV is challenging with current cell lines like Vero.
• Vero cells have limitations in achieving high cell densities for efficient viral vector production.

Purpose of the Study:

• To evaluate EB66 cells as a producer cell line for oncolytic Newcastle disease virus (NDV).
• To optimize NDV production in EB66 cells using batch and semi-perfusion culture modes.
• To assess the cell growth, virus yield, and in vivo efficacy of NDV produced in EB66 cells.

Main Methods:

• Cultivation of EB66 cells in batch and semi-perfusion modes.
• Optimization of infection parameters: temperature, protease concentration (TrypLE), and multiplicity of infection.
• Production of NDV LaSota strain in stirred tank and orbital shaken bioreactors.
• Evaluation of virus potency and in vivo tumor reduction in a mouse model.

Main Results:

• EB66 cells demonstrated effective cell growth and high cell-specific virus productivity.
• Batch processes achieved up to 4.2 × 10^8 TCID50/mL NDV with a yield of 41 TCID50/cell.
• Semi-perfusion runs reached 6.5 × 10^7 cells/mL and a virus titer of 7.5 × 10^8 TCID50/mL.
• Produced NDV showed potency and reduced tumor size in mice.

Conclusions:

• EB66 cells are a highly effective host cell line for the large-scale production of oncolytic NDV.
• Process optimization in EB66 cells enables enhanced viral vector yields.
• EB66-derived NDV holds potential for effective cancer therapy.