Clinical-Grade Oncolytic Adenovirus Purification Using Polysorbate 20 as an Alternative for Cell Lysis

Area of Science:

• Biotechnology
• Virology
• Chemical Engineering

Background:

• Oncolytic virus therapy, particularly using adenoviruses, is a promising cancer treatment.
• Current purification methods often use Triton X-100, a surfactant facing regulatory scrutiny.
• There is a need for safer, effective alternatives in adenovirus purification for clinical applications.

Purpose of the Study:

• To develop and evaluate a scalable purification strategy for oncolytic adenovirus.
• To assess Polysorbate 20 as a replacement for Triton X-100 in cell lysis for adenovirus purification.
• To ensure the purification process yields a product suitable for Phase I and II clinical trials.

Main Methods:

• A downstream purification train including clarification, tangential flow filtration, anion exchange chromatography, and polishing was designed.
• The process was evaluated using both Polysorbate 20 and Triton X-100 for cell lysis.
• Product recovery, infectious viral particle yield, host cell protein removal, and ds-DNA depletion were quantified.

Main Results:

• Polysorbate 20 effectively replaced Triton X-100 in cell lysis without compromising product recovery, potency, or purity.
• The overall recovery of infectious viral particles was 61 ± 4%.
• Host cell proteins and ds-DNA were depleted by 99.9% and 97.1%, respectively.

Conclusions:

• Polysorbate 20 is a viable and safe alternative surfactant for oncolytic adenovirus cell lysis and purification.
• The developed purification process is scalable and yields high-purity adenovirus for clinical use.
• This advancement supports the broader application of oncolytic adenovirus therapy in cancer treatment.