Oncolytic virus scalability affinity chromatography process

Abstract

Background & Aim Immunotherapy clinical trials require high titer virus preparation to adequately deliver the therapeutic transgene to clinical subjects. However, current downstream manufacturing processes for enveloped viral vectors often face challenges with limited titer production and purification scalability due to the complicated purification strategy of the virus vector. A large shift in technology must take place to enable industrialization of this process with a reduced cost in mind. Here, we will present a successful case study for purification of an enveloped virus produced using the iCELLis 500 manufacturing-scale bioreactor. The purification challenges of an enveloped viral vector increasing overall recovery without compromising its functionality will be discussed. Methods, Results & Conclusion The purification process was developed at a small scale followed by a linear scale up to the process scale. In brief, the downstream steps included a DNA removal using Nuclease treatment in the iCELLis 500 reactor followed by clarification with the Pall Stax system using V100P/HDCII depth filters, affinity column chromatography, concentration/diafiltration with Pall Cadence TFF membranes, excipient addition, and a final step of sterile filtration. The FFU step recovery titer data for this chromatography process was 60-74%. The HCP level was reduced >95% from initial levels. Overall, our purification strategy of scaling up from 1L to 250L process scale was proved to be successful with a similar overall titer recovery and HCP reduction.