TransIT-VirusGEN® Select Transfection Reagent

Large Scale Virus Production for Preclinical and Early Phase Clinical Trials

All TransIT-VirusGEN® reagents are designed to enhance delivery of packaging and transfer vector DNA to suspension and adherent HEK 293 cell types in order to increase production of recombinant lentivirus and adeno-associated virus (AAV).

TransIT-VirusGEN® SELECT Transfection Reagent is identical in formulation to our research-grade TransIT-VirusGEN® and includes release testing and quality documentation to streamline the process of ancillary material* qualification, ensuring seamless transitions from discovery through large-scale manufacturing.

With performance that exceeds PEI and PEIpro® reagents, TransIT-VirusGEN® SELECT offers a simplified, cost-effective workflow, making it the superior choice for large-scale virus production.

*USP <1043> ANCILLARY MATERIALS FOR CELL, GENE AND TISSUE-ENGINEERED PRODUCTS

• Performance – Efficient DNA delivery for large-scale production of high-titer viral vectors
• Quality – Tested for performance, identity, sterility, endotoxin and mycoplasma
• Reliability – Exceptional lot-to-lot consistency
• Flexibility – Compatible with different virus production platforms and repeat filtration
• Animal Origin Free – Fully synthetic transfection reagent formulation

TransIT-VirusGEN® SELECT Outperforms Competitor 25 kDa linear PEI and PEIpro® Transfection Reagents in Suspension and Adherent Cell Culture Systems. Lentivirus was produced using (A) suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium or (C) adherent 293T/17 cells grown in DMEM + 10% FBS and transfected with 3rd generation vectors pLKO.1-puro-CMV-TurboGFP™ transfer vector (Sigma) and ViraSafe™ Pantropic Packaging mix (pRSV-Rev, pCMV-VSV-G, pCgpV, Cell Bio Labs) 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, vol:wt), 25 kDa linear PEI (4:1, Polysciences) or PEIpro® (2:1, Polyplus Transfection). 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 (B) suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium or (D) adherent 293T/17 cells grown in DMEM + 10% FBS and transfected using pAAV-hrGFP, pAAV-RC, and pAAV-Helper plasmids (1:1:1 DNA ratio, 1.5 μg/ml plasmid DNA, Agilent Technologies) using TransIT-VirusGEN® SELECT Transfection Reagent (2:1, vol:wt), 25 kDa linear PEI (2:1, Polysciences) or PEIpro® (2:1 or 4:1, Polyplus Transfection). 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.

Lot-to-Lot Consistency with the TransIT-VirusGEN® SELECT Transfection Reagent.
(A) Lentivirus was produced using suspension FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium and transfected with 3rd generation vectors pLKO.1-puro-CMV-TurboGFP™ transfer vector (Sigma) and ViraSafe™ Pantropic Packaging mix (pRSV-Rev, pCMV-VSV-G, pCgpV, Cell Bio Labs) at a 3:0.5:0.5:2 DNA ratio, 1 ug/ml total plasmid, using the TransIT-VirusGEN® SELECT Transfection Reagent (3:1, vol:wt). 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 FreeStyle™ 293-F cells grown in FreeStyle™ F17 Medium and transfected using pAAV-hrGFP, pAAV-RC, and pAAV-Helper plasmids (1:1:1 DNA ratio, 1.5 µg/ml, Agilent Technologies) using TransIT-VirusGEN® SELECT Transfection Reagent (2:1, vol:wt). 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.