The challenge of optimizing AAV vector production
Adeno-associated virus (AAV) vectors have become a cornerstone of modern gene therapy due to their efficiency and safety. However, the industry still faces a significant manufacturing bottleneck: scaling up production while maintaining high yields, quality, and consistency.
HEK293 cell-based production systems, although widely used, can be variable and challenging to scale. Low proportions of genome-containing particles, batch-to-batch inconsistency, and the reliance on animal-derived media components further complicate production. To overcome these challenges, optimized and animal-free supplements are essential for reproducibility and regulatory compliance.
Enhancing AAV production with Recombinant Insulin
Recombinant Insulin is a well-established, animal-free media supplement that supports cellular metabolism and productivity. We investigated its potential to improve AAV2 vector production in HEK293 suspension cultures by assessing how adding insulin before transfection affects yield, encapsidation, and product quality.
Our application note demonstrates that adding Recombinant Insulin two hours before transfection significantly enhanced both capsid protein (CP) and vector genome (VG) titers, with the optimal results achieved at concentrations of 10–20 mg/L. Across multiple production runs, insulin supplementation led to:
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Higher AAV2 titers, increasing volumetric productivity.
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Improved encapsidation efficiency, with a higher proportion of full, genome-containing capsids.
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Maintained product quality, with unchanged VP1/VP2/VP3 profiles and >99% monomer content, ensuring stability and regulatory comparability.
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Preserved vector potency, as confirmed by GFP expression assays in HEK293T cells.
These findings demonstrate that Recombinant Insulin enhances AAV production efficiency without compromising product integrity.
Download the application note
Ready to enhance your AAV production? Download the application note to discover how Recombinant Insulin can improve yield, encapsidation, and process robustness in HEK293 suspension cultures.
