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VirusGEN GMP LV Transfection Kit

Proven Nucleic Acid Delivery for Large-Scale Virus Production

SKU MIR 6825-GMP Categories , ,

Description

TransIT-VirusGEN® GMP Transfection Reagent is 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). Key benefits of TransIT-VirusGEN® GMP Transfection Reagent include:

  • Performance – Efficient DNA delivery for large-scale production of high-titer viral vectors
  • Quality – GMP manufactured and tested for appearance, identity, sterility, endotoxin and mycoplasma
  • Flexibility – Compatible with different virus production platforms and repeat filtration
  • Animal Origin Free – Fully synthetic transfection reagent formulation
With industry-leading performance in high-titer virus manufacturing, TransIT-VirusGEN® GMP offers a simplified, cost-effective workflow, making it the superior choice for large-scale therapeutic virus production. Titers can be further increased with optimized enhancers included in the VirusGEN® AAV Transfection Kit and VirusGEN® GMP LV Transfection Kit.

For information on identity and residual reagent assays available through partner contract service laboratories, please contact techsupport@mirusbio.com.

Example Certificate of Analysis (PDF)

Example Certificate of Origin and BSE TSE Statement (PDF)

TransIT-VirusGEN® GMP is intended to be used as an ancillary material for cell, gene and tissue-engineered products, per USP <1043>.

SKU: MIR 6825-GMP

 

 

Supporting Data

TransIT-VirusGEN® Reagent and the VirusGEN® LV Transfection Kit Outperform PEI-based Reagents with Multiple Lentivirus Packaging Systems. Lentivirus production using TransIT-VirusGEN® Transfection Reagent +/- the VirusGEN® LV Enhancer was compared to Competitor A3 (2:1 reagent-to-DNA ratio) and Competitor C1 (4:1 reagent-to-DNA ratio) using their recommended protocol. Lentivirus was produced by transfecting Expi293™ cells grown in Expi293™ Expression Media (Thermo Fisher Scientific) at 4 x 106 cells/ml with two different LV vector mixtures (1 μg/ml, 2μg/ml) including: (A) 3rd generation vectors pALD-LentiEGFP-A transfer vector and pALD-VSV-G-A, pALD-Rev-A, pALD-GagPol-A packaging vectors (3:0.5:0.5:2 DNA ratio, Aldevron) or (B) 3rd generation vectors pLKO.1-puro-CMV-TurboGFP™ transfer vector (MilliporeSigma) and ViraSafe™ Pantropic Packaging mix (pRSV-Rev, pCMV-VSV-G, pCgpV, CellBio Labs) at a 3:0.5:0.5:2 DNA ratio. The VirusGEN® LV Enhancer was added to the TransIT-VirusGEN® condition at 18 hours post-transfection. Virus containing supernatant was used to transduce 293T/17 cells and GFP expression was measured at 72 hours post-transduction using Guava® easyCyte™ 5HT Flow Cytometer. Lentivirus functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the range of duplicate wells.

TransIT-VirusGEN® Reagent and VirusGEN® LV Transfection Kit are Compatible with Multiple Cell Lines and Media Formulations for LV Production in Suspension 293 Cells. Viral Production cells (293-VP) grown in LV-MAX™ Production Medium (ThermoFisher), Expi293™ cells grown in Expi293™ Expression Medium (ThermoFisher), or FreeStyle™ 293-F cells grown in FreeStyle™ F17 (ThermoFisher) were seeded at 4 x 10^6 cells/ml prior to transfection (2 ml/non-treated 6-well plate). Lentivirus was produced using the TransIT-VirusGEN® Transfection Reagent +/- VirusGEN® LV Enhancer (Mirus Bio, 3:1 reagent-to-DNA ratio (wt:vol)) according to the recommended protocol. Lentivirus was produced by transfecting 3rd generation vectors pALD-LentiEGFP-A transfer vector and pALD-VSV-G-A, pALD-Rev-A, pALD-GagPol-A packaging vectors at a 3:0.5:0.5:2 DNA ratio (Aldevron, 25 μg/flask (1 μg/ml final concentration). The VirusGEN® LV Enhancer was added 18 hours post-transfection. Functional titers from virus containing supernatant was measured following transduction 293T/17 cells and GFP expression was measured at 72 hours post post-transduction using Guava® easyCyte™ 5HT Flow Cytometer. Lentivirus functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the standard deviation of triplicate wells.

Resources

Specifications

Storage Conditions:
VirusGEN® GMP LV Enhancer: Store at 2 to 10°C
VirusGEN® GMP LV Complex Formation Solution: Store at 2 to 10°C
VirusGEN® GMP LV Transfection Kit: Multiple storage conditions – see individual bottles for specific recommendations
TransIT-VirusGEN® GMP Transfection Reagent: Store at -10 to -30°C

Product Guarantee:
All Configurations: Refer to Certificate of Analysis for Retest Date

Usage Statement:
All Configurations: For Research Use and Further Manufacturing; Not for Administration into Humans.

Animal Origin Statement:
All Configurations: This product is animal origin free.

Technical Product Literature

Full Protocols
TransIT-VirusGEN® GMP Protocol (PDF)
VirusGEN® GMP LV Transfection Kit Protocol (PDF)

SDS
TransIT-VirusGEN® GMP SDS (PDF)
VirusGEN® GMP LV Complex Formation Solution SDS (PDF)
VirusGEN® GMP LV Enhancer SDS (PDF)
VirusGEN® GMP LV Transfection Kit SDS (PDF)

Additional Resources

For a lot-specific Certificate of Analysis (COA) or Certificate of Origin (COO) containing a TSE/BSE Statement for this product, please contact techsupport@mirusbio.com.

FAQs

See the TransIT-VirusGEN GMP FAQs

See the VirusGEN LV FAQs

Using transfection reagents and enhancers from Mirus has given our platform a competitive advantage. Increasing efficiency and lowering costs for all of our porgrams.

 

Sally Mader, PhD
ZYZ Theraputics