Large-scale AAV production: TransIT-VirusGEN SELECT

TransIT-VirusGEN® SELECT from Mirus Bio is a high-performance transfection reagent for the large-scale production of viral vectors. With exceptional lot-to-lot consistency and having undergone extensive quality release testing, this reagent is ideal for the production of high-titre viral vectors for preclinical and early phase clinical trials.

Titers can be further increased with optimised enhancers included in the TransIT-VirusGen SELECT AAV Transfection Kit and TransIT-VirusGen SELECT LV Transfection Kit.

Benefits of using TransIT-VirusGEN SELECT
• Highly efficient DNA delivery
• Ideal for large-scale production of viral vectors
• Animal origin free
• Exceptional lot-to-lot consistency
• Compatible with a variety of recombinant virus platforms & cell culture systems
• Extensively characterised to confirm formulation identity, sterility, low endotoxin levels & absence of mycoplasma

TransIT-VirusGEN SELECT features

Read more about the research grade alternative, TransIT-VirusGEN, which is ideal for research and development.

TransIT-VirusGEN SELECT outperforms competitor 25 kDa linear PEI & PEIpro® transfection reagents in suspension & adherent cell culture systems

Lentivirus was produced using suspension (A) or adherent (C) cells grown transfected with 3rd generation vectors pLKO.1-puro-CMV-TurboGFP™ transfer vector and ViraSafe™ pantropic packaging mix at a 3:0.5:0.5:2 DNA ratio, 1 μg/ml total plasmid DNA, using the TransIT-VirusGEN SELECT transfection reagent (3:1), 25 kDa linear PEI (4:1) or PEIpro® (2:1). Virus-containing supernatant was used to transduce 293T/17 cells and GFP expression was measured at 72 hours post-transduction using a Guava® easyCyte™ 5HT Flow Cytometer.

AAV2 was produced using suspension (B) or adherent (D) cells transfected using pAAV-hrGFP, pAAV-RC, and pAAV-Helper plasmids (1:1:1 DNA ratio) using TransIT-VirusGEN SELECT Transfection Reagent (2:1), 25 kDa linear PEI (2:1) or PEIpro® (2:1 or 4:1). Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction using a Guava® easyCyte™ 5HT Flow Cytometer. For both lentivirus and AAV, functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the range of triplicate wells.

TransIT-VirusGEN SELECT has exceptional lot-to-lot consistency

(A) Lentivirus was produced using suspension cells grown transfected with 3rd generation vectors pLKO.1-puro-CMV-TurboGFP™ transfer vector and ViraSafe™ Pantropic Packaging mix at a 3:0.5:0.5:2 DNA ratio, 1 ug/ml total plasmid, using the TransIT-VirusGEN SELECT Transfection Reagent (3:1). Virus containing supernatant was used to transduce 293T/17 cells and GFP expression was measured at 72 hours post-transduction using a Guava® easyCyte™ 5HT Flow Cytometer.

(B) AAV2 was produced using suspension transfected using pAAV-hrGFP, pAAV-RC, and pAAV-Helper plasmids (1:1:1 DNA ratio) using TransIT-VirusGEN SELECT Transfection Reagent (2:1). Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction using a Guava® easyCyte™ 5HT Flow Cytometer. For both lentivirus and AAV, functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the standard deviation of triplicate wells.

Applications
• Large-scale virus production

Material available for download
TransIT-VirusGEN SELECT full protocol
TransIT-VirusGEN SELECT example certificate of analysis
TransIT-VirusGEN SELECT example certificate of origin

PEIpro® is a trademark of Polyplus-Transfection
TurboGFP™  is a trademark of Evrogen, JSC
Guava® and EasyCyte™are registered trademarks of Guava Technologies, Inc.