TransIT AAViator™ GMP Transfection Reagent

Supporting Data

AAV9 GFP Transgene Vector Production

AAV8 GFP Transgene Vector Production

AAV5 GFP Transgene Vector Production

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.

AAV9 RPE65 Transgene Vector Production

AAV8 RPE65 Transgene Vector Production

AAV5 RPE65 Transgene Vector Production

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

Plasmid DNA Dose

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.