Achieve Peak AAV Titers with the AAViator™ System
Stop compromising between AAV titer, quality, and cost. Manufacturing high-yield, high-quality adeno-associated virus for gene therapy presents significant challenges, from low particle yield and high percentages of empty capsids to the prohibitive cost of GMP-grade plasmid DNA.
The TransIT®-AAViator™ Transfection System from Mirus Bio is a complete, two-component solution engineered to overcome these hurdles. This chemically-defined system combines the powerful TransIT®-AAViator Transfection Reagent with the synergistic RevIT™ AAV Enhancer to deliver exceptional, industry-leading AAV titers in suspension HEK 293 cell systems.
Key Benefits:
- Exceptional Viral Titers: Achieve market-leading functional AAV yields in large-scale suspension cultures.
- Superior Capsid Quality: Significantly increases the percentage of full capsids, enhancing the potency of your final product and simplifying downstream processing.
- Drastically Reduce Costs: The highly efficient system allows for a reduction in plasmid DNA usage by up to 50% compared to competitor reagents (like PEI), slashing raw material costs.
- Seamless Scalability: Robust, reproducible performance that scales from small-scale shake flasks to large-scale stirred-tank bioreactors.
- Chemically Defined: Fully synthetic, animal-origin-free (AOF) components for a clear regulatory path.
Supporting Data
TransIT-AAViator System Outperforms Competitor Reagents in Suspension AAV Cell Cultures with a GFP transgene
Viral Production 2.0 cells grown in Viral Production Medium were transfected with pAAV-WPRE-GFP (Aldevron), pAAV-Helper (Aldevron) and pAAV-RepCap (AAV9 and AAV8 Genemedi, AAV5 Aldevron) at a 1:1:1 plasmid concentration ratio with the following reagents: TransIT-AAViator® Transfection reagent (1.25:1,vol:wt), Polymer F (1:1) , or Polymer P (1:1). RevIT-AAV Enhancer (1 µl/mL) was added at the time of transfection. All AAV serotypes were harvested at 72 hours post-transfection using chemical lysis. Genome titers were determined via dPCR using primers and a probe targeting the CMV promoter region. Total assembled capsids were determined using the AAV9, AAV8 or AAV5 Titration ELISA Kits (Progen), and the Percent Full Capsids were determined by dividing the number of genome copies by total assembled capsids for each condition. The error bars represent the standard deviation of duplicate wells.
TransIT-AAViator System Outperforms Competitor Reagents in Suspension AAV Cell Cultures with an RPE65 transgene
Viral Production 2.0 cells grown in Viral Production Medium were were transfected with pAAV-WPRE-RPE65 (Aldevron), pAAV-Helper (Aldevron) and pAAV-RepCap (AAV9 and AAV8 Genemedi, AAV5 Aldevron) at a 1:1:1 plasmid concentration ratio with the following reagents: TransIT-AAViator® Transfection reagent (1.25:1,vol:wt), Polymer F (1:1) , or Polymer P (1:1). RevIT-AAV Enhancer (1 µl/mL) was added at the time of transfection. All AAV serotypes were harvested at 72 hours post-transfection using chemical lysis. Genome titers were determined via dPCR using primers and a probe targeting the CMV promoter region. Total assembled capsids were determined using the AAV9, AAV8, or AAV5 Titration ELISA Kits (Progen), and the Percent Full Capsids were determined by dividing the number of genome copies by total assembled capsids for each condition. The error bars represent the standard deviation of duplicate wells.
High AAV Titers Across A Range of Plasmid DNA Doses
Viral Production 2.0 cells grown in Viral Production Medium were were transfected with pAAV-WPRE-GFP (Aldevron), pAAV-Helper (Aldevron) and pAAV-RepCap (AAV 9 Genemedi) at a 1:1:1 plasmid concentration ratio with TransIT-AAViator® Transfection reagent (1.25:1,vol:wt). RevIT-AAV Enhancer (1 µl/mL) was added at the time of transfection. AAV was harvested at 72 hours post-transfection using chemical lysis. Genome titers were determined via dPCR using primers and a probe targeting the CMV promoter region. The error bars represent the standard deviation of duplicate wells.
Specifications
- Storage Conditions: Store TransIT®-AAViator Reagent at 4°C. Store RevIT™ AAV Enhancer at -20°C (protect from light).
- Product Guarantee: Guaranteed for 6 months from the date of shipment when stored properly.
- Usage Statement: For Research Use Only. Not for use in diagnostic or therapeutic procedures.
- Animal Origin Statement: This product is chemically defined and animal-origin-free.
- This Product Requires Dry Ice Shipment (for the RevIT™ AAV Enhancer component).