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TransIT-VirusGEN® SELECT Transfection Reagent and Kits

Large Scale Virus Production for Preclinical and Early Phase Clinical Trials

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MIR 673030 ml - for 10 L of cell cultureMIR 6735New150 ml - for 50 L of cell culture

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TransIT-VirusGEN® SELECT 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® SELECT Transfection Reagent include:

  • Performance – Efficient DNA delivery for large-scale production of high-titer viral vectors
  • Quality – Tested for performance, 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® SELECT offers a simplified, cost-effective workflow, making it an excellent choice for large-scale virus production in preclinical studies and early-phase clinical trials. Titers can be further increased with optimized enhancers included in the VirusGEN® SELECT AAV Transfection Kit and VirusGEN® SELECT LV Transfection Kit. When transitioning to late-phase clinical trials or commercial manufacturing, we recommend TransIT-VirusGEN® GMP.

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Example Certificate of Analysis

Example BSE TSE Statement

 

Compare TransIT-VirusGEN® Transfection Reagents

Therapeutic Development Pipeline
Research & Development
Preclinical & Early-phase Clinical Trial
Late-phase Clinical Trial & Commercial Manufacturing
Composition
Identical TransIT-VirusGEN® Reagent Formulation
Ready to use, chemically defined, animal origin free in 77.5% ethanol
LV and AAV Enhancer Kits Available
 
Coming Soon
 
Usage
R&D
Preclinical
GMP
Quality Control
Functional AAV titer assay
Functional AAV titer assay
Formulation Identity
Appearance
Sterility per USP <71>
Bacterial endotoxin per USP <85>
Mycoplasma per USP <63>
Validated Formulation Identity Assay
Appearance
Sterility per USP <71>
Bacterial endotoxin per USP <85>
Mycoplasma per USP <63>
Configuration and Packaging
0.3 ml vial
0.75 ml vial
1.5 ml vial
5 x 1.5 ml vials
10 x 1.5 ml vials
30 ml bottle
150 ml bottle
150 ml bottle
Manufacturing
Research grade manufacturing with aseptic filtration
Research grade manufacturing with sterile filtration
GMP validated manufacturing process with sterile filtration
Raw Materials
Research grade
Research grade
GMP-grade critical raw materials
Available documentation
Certificate of Analysis
Certificate of Origin
Certificate of Analysis
Certificate of Origin
TSE/BSE Statement
Certificate of Analysis
Certificate of Origin with TSE/BSE Statement
DMF Available in 2022 with Quality Agreement
Available Analytical Assays
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Validated Identity Assay
Residual Reagent Assay
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Figures and Data

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TransIT-VirusGEN and the VirusGEN AAV Transfection Kit Produce High AAV Titers at the 1L Scale

TransIT-VirusGEN® Reagent and the VirusGEN® AAV Transfection Kit Generate Higher AAV Titers. (A) Expi293F™ cells (Thermo Fisher) grown in Expi293™ Expression Media (Thermo Fisher) were used to generate recombinant AAV2 via transient transfection using TransIT-VirusGEN® Transfection reagent (1.5:1 reagent-to-DNA ratio (vol:wt), Mirus Bio LLC), VirusGEN® AAV Transfection Kit (Mirus Bio LLC), PEIpro® (1:1, Polyplus) or FectoVIR®-AAV (1:1, Polyplus) using their recommended protocol.  AAV2 was produced by transfecting pAAV-hrGFP, pAAV-RC and pAAV-Helper plasmids (1:1:1 DNA ratio, 2 μg/ml-2 mg per flask, Agilent Technologies). Cells were transfected at a cell density of 2 x 106 cells/ml, 1 L in a 2.8 L Thomson shake flask. AAV2 was harvested at 72 hours post-transfection using chemical lysis. (A) Functional titers were determined via transduction of HT-1080 cells and GFP expression was measured 48 hours post-transduction using Guava® easyCyte™ 5HT Flow Cytometer. AAV functional titers were measured from virus dilutions with less than 20% GFP positive cells. Genome copies were determined by ddPCR using primers and a probe targeting the CMV promoter. (B) Total assembled capsids were determined using the AAV2 Titration ELISA Kit (Progen) and the GC/capsid ratio was determined by dividing the number of genome copies by total assembled capsids for each condition. The error bars represent the range of duplicate flasks.

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VirusGEN-GMP-AAV5683_5702_AAV media adaptations

TransIT-VirusGEN® Reagent and the VirusGEN® AAV Transfection Kit Outperform PEI-based Reagents for AAV Production across Different Suspension Cell Types and Media Formulations. (A) Expi293F™ cells (Thermofisher) or (B) Viral Production cells (Thermofisher) were grown or adapted to Expi293™ Media (ThermoFisher), LV-MAX Production (ThermoFisher), FreeStyle™ F17 (ThermoFisher), or BalanCD HEK293 (Irvine Scientific) media prior to transfection. AAV production using TransIT-VirusGEN® Transfection reagent (2:1 reagent-to-DNA ratio (vol:wt), Mirus Bio LLC), VirusGEN® AAV Transfection Kit (Mirus Bio LLC) was compared to PEIpro® (1:1 reagent-to-DNA ratio (vol:wt), Polyplus) and PEI MAX (2:1 reagent-to-DNA ratio (vol:wt), Polysciences) using their recommended protocol. AAV2 was produced by transfecting pAAV-hrGFP, pAAV-RC, and pAAV-Helper plasmids (1:1:1 DNA ratio, 1.5 μg/ml = 3 μg/well, Agilent Technologies). Cells were transfected at a cell density of 2 million cells/ml. Harvested virus was used to transduce HT1080 cells and GFP expression was measured 48 hours post-transduction using Guava® easyCyte™ 5HT Flow Cytometer. AAV functional titers were measured from virus dilutions with less than 20% GFP positive cells. The error bars represent the range of duplicate wells.

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VirusGEN-GMP-Lentivirus-5531_LV Vector_Competitor

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 PEIpro® (2:1 reagent-to-DNA ratio, Polyplus) and PEI MAX (4:1 reagent-to-DNA ratio, Polysciences) using their recommended protocol. Lentivirus was produced by transfecting Expi293™ cells grown in Expi293™ Expression Media (Thermo Fisher) 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.

 
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VirusGEN-GMP-Lentivirus-5566_LV system_to_system

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.

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TransIT-VirusGEN SELECT Multiple Filtrations

TransIT-VirusGEN® Transfection Reagent can be Filtered Multiple Times with No Effect on Reagent Performance. 
(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® Transfection Reagent (3:1, vol:wt) that was filtered through a 0.22 um polyethersulfone (PES) filter unit (Millipore Sigma) for the indicated number of times. 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® 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.

 

Resources

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

Specifications

Storage Conditions:
TransIT-VirusGEN® SELECT Transfection Reagent: Store at -10 to -30°C
VirusGEN® SELECT AAV Transfection Kit: Multiple storage conditions - see individual bottles for specific recommendations
VirusGEN® SELECT LV Transfection Kit: Multiple storage conditions - see individual bottles for specific recommendations

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

Technical Product Literature


Additional Resources

Testimonials

Mirus sent me samples of TransIT-VirusGEN® to test with 293T cells to generate lentivirus. It outperforms the transfection reagent I was using before. The protocol is really simple, and the transfections are robust. I’m getting great virus production.
Dr. Steve Williams
Xyphos Inc.
I am definitively getting better transfection efficiency with TransIT-VirusGEN® but more important, a higher level of transduced cells afterwards. Of course, efficiency depends on backbone and transgene features, but I am very satisfied.
Dr. Zuzana Drobna
North Carolina State University