Additional Insights
A robust and scalable upstream process is the foundation for manufacturing advanced therapeutics. For complex biologics like AAV vectors, moving from research-scale adherent cultures to a large-scale suspension platform requires a systematic, data-driven approach. Applying proven intelligence in vector production accelerates the path to generating sufficient material for IND-enabling toxicology studies and subsequent clinical evaluation.
The selection of the production platform itself is a primary determinant of vector quality. Data indicates that the choice of expression system : for example, mammalian HEK293 versus insect Sf9 cells : can result in significant differences in capsid properties and biological potency (PMID: 37597192). A well-characterized mammalian-based suspension process provides a more direct route to a clinically relevant product profile, minimizing risks associated with vector efficacy.
Our upstream process development focuses on creating a reproducible and scalable manufacturing solution tailored to a specific AAV serotype and transgene. This involves a multi-parameter optimization strategy to maximize productivity while maintaining vector integrity.
Key areas of focus include:
- Media Selection and Feed Strategy: Evaluating and selecting optimal basal media and feed supplements to support high-density cell growth and extend the production phase in single-use bioreactors.
- Transfection Parameter Optimization: Systematically refining the ratio of helper, rep/cap, and transgene plasmids, along with the transfection reagent, to achieve maximum vector yield per cell.
- Bioreactor Parameter Control: Establishing precise control strategies for pH, dissolved oxygen (DO), and temperature to ensure consistent performance from 2L bench-scale runs to 500L+ production volumes.
- Scale-Up Confirmation: Performing confirmation runs at increasing volumes to verify that process parameters translate effectively, ensuring predictable performance and product quality at scale.
This methodical development work provides the process knowledge continuity required for a successful technology transfer into a GMP environment. By defining a robust upstream process early, programs can avoid costly delays and generate the high-quality vector needed to meet aggressive clinical timelines.
Return to the main Large-Scale AAV Manufacturing and Process Development hub to explore our downstream and analytical capabilities.
This site tour showcases Franklin Biolabs’ preclinical and bioanalytical facilities, spanning over 100,000 square feet of animal housing and specialized laboratory space.
