Avoiding Scale-Up Pitfalls
Upstream Process Development Strategies for AAV Manufacturing
How to optimize transfection efficiency, plasmid ratios, and process scale-up to maximize AAV yield and percent full capsids
Avoiding AAV Scale-Up Pitfalls
Upstream Process Development Strategies for AAV Manufacturing
How to optimize transfection efficiency, plasmid ratios, and process scale-up to maximize AAV yield and percent full capsids in a 200 L bioreactor
Your Trail Guide from Benchtop to Bioreactor
Scaling AAV transfection isn’t just about getting bigger – it’s about getting better while avoiding risks that can kill your drug asset. This webinar covers the critical process optimizations and side-steps that make the difference between success and failure at large scale.
Key Takeaways:
How to optimize transfection for higher titers and more full capsids
The #1 mistake that leads to poor yield and how to avoid it
Real-world data on TransIT-VirusGEN® and RevIT™ in large-scale bioreactors
A case study on how process refinements improved AAV production
30 min
Experts, Sandy Tseng and Hugh Murray discuss challenges and solutions when scaling AAV manufacturing.
Your Trail Guide from Benchtop to Bioreactor
30 min
Scaling AAV transfection isn’t just about getting bigger – it’s about getting better while avoiding risks that can kill your drug asset. This webinar covers the critical process optimizations and side-steps that make the difference between success and failure at large scale.
Key Takeaways:
How to optimize transfection for higher titers and more full capsids
The #1 mistake that leads to poor yield and how to avoid it
Real-world data on TransIT-VirusGEN® and RevIT™ in large-scale bioreactors
A case study on how process refinements improved AAV production
Pitfall Previews
Pitfall
Inefficient Mixing
Problem: Poor homogenization when adding transfection complexes to bioreactors leads to uneven uptake and lower titers.
Solution: Adjusting the addition point can significantly improve complex distribution and efficiency.
“We saw that where we added the complex to the bioreactor made a big difference. Instead of adding it to the top, we added it to the bottom, shooting it right into the impeller. TransIT-VirusGEN is very stable so there was no problem with the shear. That gave us better mixing and a more homogeneous distribution of the complex.”
Process Development Senior Scientist
MilliporeSigma
Pitfall
Oversized Transfection Complexes
Problem: Complexes that are too large fail to transfect efficiently, reducing productivity.
Solution: Modify the volume of your complex and the concentration of the DNA within it while using the best transfection reagent and optimize via Dynamic Light Scattering (DLS) tests to ensure complexes remain within the optimal size range for better transfection efficiency.
Pitfall
Unoptimized Plasmid Ratio
Problem: Incorrect plasmid DNA ratios lead to inefficient transfection and lower vector production.
Solution: Optimizing plasmid ratios ensures the right balance for high titers and consistent capsid packaging.
MilliporeSigma Optimization Process:
Optimize the makeup of total DNA between your three plasmids: the pGOI, the pHelper, and the pRepCap
Pitfall
Inaccurate Scale-Up Estimates
“That’s the nightmare scenario:
you do all this work, everything looks good, you scale it up, and you get half the yield. That’s a that’s a bad look and clients do not like that.“
Process Development Senior Scientist
MilliporeSigma
Problem: When scaling from bench-scale to large bioreactors, CDMOs must ensure that projected yields remain consistent at GMP scale. Yield losses can be significant, leading to dissatisfied clients and costly setbacks.
Solution: Carefully controlling process variables like heat transfer, aeration, gassing, and mixing at each scale ensures reliable, reproducible results. By integrating real-time monitoring and scaling studies, manufacturers can confidently predict performance at full production scale.
Pitfall
Complexation Issue Detection
Problem: Without real-time measurement tools, it’s difficult to detect issues with complexation that could affect transfection efficiency.
Solution: Satellite vessels and Dynamic Light Scattering (DLS) provide real-time data to ensure optimal complex size.
Pitfall
Avoid Misleading Data
Problem: Different analytical methods give different insights. Using the wrong data set could misinform your downstream decisions.
Solution: Using mass photometry and full:empty ratio analysis for the right stage of development ensures accurate data and better decision making.
Get the full solutions and webinar results
- Extended Q+A Session Access
- 10+ Avoidable AAV Pitfalls and Guides Around Them
- Product Launch Preview