Oncolytic virus purification with periodic counter-current chromatography

Area of Science:

• Biopharmaceutical manufacturing
• Advanced chromatography techniques
• Oncolytic virotherapy

Background:

• Virus-based biologicals, including oncolytic viruses, are crucial for modern medicine.
• Manufacturing scales for oncolytic viruses vary significantly, from research to commercial production.
• Periodic counter-current chromatography (PCC) presents a scalable solution to reduce equipment size and streamline purification processes.

Purpose of the Study:

• To design and evaluate an efficient PCC process for the intermediate purification of oncolytic adenovirus.
• To assess the impact of feedstock variations, post-load washing, and extended operation on virus recovery and purity.
• To demonstrate the stability and automation potential of PCC for adenovirus purification.

Main Methods:

• Development of a four-column ion-exchange PCC setup for adenovirus purification.
• Testing of three scenarios: feedstock variation, inclusion of a post-load wash step, and long-term operational stability.
• Analysis of virus recovery rates and reduction of DNA and protein impurities.

Main Results:

• Achieved virus recoveries ranging from 57% to 86%, comparable to or exceeding batch purification methods.
• Demonstrated significant impurity reduction, with over 80% DNA removal and over 70% total protein removal.
• Confirmed process stability, with effective suppression of inlet perturbations and potential for unattended operation.

Conclusions:

• PCC is a highly effective and scalable method for the intermediate purification of oncolytic adenovirus.
• The developed PCC process offers robust performance, high recovery, and excellent impurity removal.
• The automation and stability of PCC make it a promising technology for industrial-scale biopharmaceutical production.