Scaling up an AAV production platform to support clinical and commercial needs: challenges and opportunities

Transient transfection of a suspension HEK293 bioreactor is becoming the preferred method of rAAV manufacturing. This is because of its versatility to produce a wide range of constructs that are more potent than other production strategies and can enable organizations to rapidly scale up manufacturing. With all its benefits, significant challenges face manufacturers to successfully scale up and execute large scale transient transfection. At Oxford Biomedica Solutions, we have solved these challenges by investing heavily into our platform and developing techniques that allow seamless production from 2L to 2000L scale using transient transfection. In this discussion, we will show data demonstrating that through understanding of the fundamental kinetics and optimization of transfection mixture, precise equipment engineering and the development of a robust purification process we can successfully execute large scale, straightforward transient transfection up to 2000L scale. Using our approach, we are now projecting to surpass greater than 1E17 total vgs from a single 500L. This level of productivity can help drug developers fully support clinical and commercial needs and open up the gene therapy space to address larger patient populations.Equipment engineering and process control are crucial for successful scale up of transient transfection

• Transition from ultracentrifugation to anion exchange chromatography and scaling up of anion exchange chromatography to achieve over 90% full capsids
• Comparability of titer, empty to full capsid ratio and product quality between 2 and 2000-L scale
• Rational design of satellite runs to support scale-up understanding

Ify Iwuchukwu

Director of the Pilot Group at Oxford Biomedica

Dr. Ify Iwuchukwu is the Director of the Pilot Group at Oxford Biomedica (OXB) Solutions and is responsible for scaling up processes, driving technology innovation, supporting platform development, and delivering pre-clinical supply.  Prior to Oxford Biomedica Solutions, Ify worked at Joule Unlimited and GreenLight Biosciences in roles of increasing responsibility, where she led the development, scale up and technology transfer of Upstream and Downstream processes for biofuels, biopesticides, small molecules and nucleic acids.
Ify holds a BS and MSc in Chemical Engineering and a PhD in Chemical and Biomolecular Engineering from the University of Tennessee.

Taylor Cunningham

Field Application Scientist, Cytiva

Field Application Scientist with Cytiva, where he acts as a subject matter expert on single-use upstream technologies. He holds a degree in Biology and has worked on various mAb and viral vector platforms throughout his career. Prior to joining Cytiva, Taylor worked at Homology (now Oxford Biomedica) where he was an integral team member focused on the successful development and scale-up of their AAV platform

Alejandro Becerra

Principal Applications Scientist and Global Purification Technical Lead at Thermo Fisher Scientific

Principal Applications Scientist and Global Purification Technical Lead at Thermo Fisher Scientific. Alejandro is a chemical engineer by training and holds a Ph.D. from Cornell University. Dr. Becerra has over 15 years of experience in downstream processing and customer support having spent 11 years at Millipore in various bioprocess engineering roles prior to joining Thermo Fisher in 2018. Alejandro is a subject matter expert in preparative chromatography with expertise in the development, optimization and scale-up of purification processes for antibodies, recombinant proteins, viral vectors, and nucleic acids.