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Invitrogen™ LV-MAX™ Transfection Kit

Catalog No. A35348
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Catalog No. A35348 Supplier Invitrogen™ Supplier No. A35348
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Description

TheLV- MAX Transfection Kit is an essential component of the LV-MAX Lentiviral Production System.

The Gibco LV-MAX Transfection Kit is a vital component of the LV-MAX Lentiviral Production System, specifically designed for the transient transfection of high-density suspension-adapted Viral Production Cells (human embryonic kidney (HEK) 293-derived cells). This kit is designed to maximize lentiviral particle production, ensuring high yields and reproducibility.

Key features of the LV-MAX Transfection Kit

  • High LV yields—achieves functional titers up to 1 x 108 TU/mL (unconcentrated)
  • High-density growth—supports growth of Viral Production Cells at densities greater than 1 x 107 cells/mL for maximum vector output
  • AOF formulation—chemically defined, serum-free, and animal-origin-free medium ensures consistent performance by eliminating variability from undefined components
  • Ready-to-use—no media supplementation required; simplifies workflows and provides reliable, lot-to-lot reproducibility in lentivirus production
  • Expression system compatible—fully compatible with the LV-MAX Lentiviral Production System for seamless integration into end-to-end lentivirus manufacturing processes
  • Scalable and reproducible transfection and production—validated from 96–well plates to multi‐liter bioreactors, enabling easy scale-up without sacrificing cell yields.
  • Easy and robust culture and transfection protocols, seamlessly integrating into any transient production workflow

Kit components

  • LV-MAX Transfection Reagent—a high-efficiency, cationic, lipid-based reagent uniquely formulated for co-transfecting multiple plasmids into high-density Viral Production Cell cultures. This reagent ensures high transfection efficiency with minimal toxicity.
  • LV-MAX Supplement—an optimized, chemically defined, serum-free, protein-free, animal origin-free formulation designed to suppress cell growth during transfection. It enhances lentiviral vector production without compromising cell viability or interfering with the production process.
  • LV-MAX Enhancer—a proprietary, animal origin-free formulation that significantly boosts lentiviral vector production

The LV-MAX Transfection Kit is designed to deliver superior performance in lentiviral production workflows, providing researchers with reliable and scalable transfection solutions.

Applications

  • Gene therapy—ideal for the production of lentiviral vectors for gene therapy research and development
  • Cell therapy—supports the generation of lentiviral vectors for cell therapy applications, facilitating seamless transition from bench to clinic
  • Research and development—provides a reliable and scalable solution for viral vector production in various research settings

Available in RUO or CTS

Offered as both a Research Use Only version and a GMP-compliant Cell Therapy Systems (CTS) version (Cat. No. A4132602) to support translational and clinical-scale applications.

Specifications

Cell Line Established Cell Lines
Classification Animal Origin-Free, Chemically Defined, Protein-Free, Serum-Free
Content And Storage • 50 mL LV-MAX Supplement
• 6 mL LV-MAX Transfection Reagent
• 40 mL LV-MAX Enhancer

Sufficient for 1 L LV production 

Store at 2–8°C, protect from light.
Cell Type 293-derived Cells
Sample Type Plasmid DNA
Transfection Technique Lipid-based Transfection
For Use With (Application) Transfection
Product Line LV-MAX
Product Type Transfection Kit
Quantity 1 L
Serum Compatible Yes
Shipping Condition Wet Ice
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Frequently Asked Questions (FAQs)

What are the major differences between LV-MAX and CTS LV- MAX products?

The LV-MAX and CTS LV- MAX products have equivalent performance. In contrast to the research use only LV-MAX products, CTS LV-MAX products have an additional intended use for cell and gene therapy applications, and are complemented with a more robust regulatory support package to enable clinical therapeutic applications. This means that to the best of our ability, we keep our CTS product specifications and release requirements up-to-date in accordance with the latest regulatory guidance specific for this intended use.
What is the intended use claim for your LV-MAX and CTS LV-MAX products?

Our LV-MAX products are intended for research use only while our CTS LV- MAX products are intended for research use or manufacture of cell, gene or tissue based products. Please refer to the individual products for further details on the intended use claim.
Do you offer a lentivirus production service?

Yes. Please provide your details here (https://www.thermofisher.com/us/en/home/global/forms/life-science/lentiviral-vector-production-get-info.html) and mention your interest in our lentivirus production services. A member of our services team will contact you shortly.
With the LV-MAX Production System, are the lentiviral particles stable in packaging media?

Lentiviral particles packaged in our LV-MAX Production Medium will be stable at 4 degrees C for 24 hours and will then begin to lose activity. For long-term storage, we recommend storing at –80 degrees C.
With the LV-MAX Production System, can I use cell lines besides HT1080 for titering my lentivirus?

We strongly recommend the human fibrosarcoma line HT1080 (ATCC, Cat. No. CCL-121) as the “gold standard” for reproducibly titering lentivirus. However, you may wish to use the same mammalian cell line to titer your lentiviral stocks as you will use to perform your expression studies (e.g., if you are performing expression studies in a dividing cell line or a non-primary cell line). If you have more than one lentiviral construct, we recommend that you titer all of the lentiviral constructs using the same mammalian cell line.
With the LV-MAX Production System, do I need to perform spinoculation for viral titering? What will happen if I don't?

Spinoculation increases lentiviral transfection efficiency and provides higher viral titer than without the spin step. If you don't spin, you can expect to get 5- to 10-fold less lentiviral titer.

What transfer plasmid:packaging vectors ratio should I use for large-insert genes?

We recommend a weight-to-weight ratio of 2:3 transfer plasmid:packaging vectors; if needed use a 3:3 weight-to-weight ratio.
With the LV-MAX Production System, should I optimize the ratios of my transfer plasmid and packaging vectors?

In the user guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0017000_LV_MAX_ViralProductionSystem_UG.pdf ), we have provided recommendations based on our development of the kit. If you need to optimize further for your specific need, we recommend a weight-to-weight ratio of transfer plasmid:packaging vectors at 1:3, 2:3, and 3:3.
With the LV-MAX Production System, how important are the incubation times in the protocol?

The critical incubation time is the one for the diluted LV-MAX Transfection Reagent. Prior to combining with diluted DNA, the diluted LV-MAX Transfection Reagent should be incubated for only 1 min, and not more than 4 mins.
Will the LV-MAX Production System work if I have SV40 elements in my vector?

Yes, that should be fine.
Are the Viral Production Cells 293T-derived?

No, these cells are derived from Freestyle 293F, and have been adapted to our LV-MAX Production Medium.
Are all components of the LV-MAX Production System animal origin-free?

Yes, all components of the LV-MAX Production System are animal origin-free. However, if Opti-MEM I Reduced Serum Medium is used as the DNA-LV-MAX Transfection Reagent complexation medium, it will no longer be animal origin-free.
Are all components of the LV-MAX Production System serum-free?

Yes, all components of the LV-MAX Production System are serum-free.
Are all components of the LV-MAX Production System GMP-certified?

In the LV-MAX Production System, only the LV-MAX Production Medium is manufactured according to cGMP guidelines. However, we do offer our CTS LV- MAX products (CTS LV-MAX Production Medium, CTS Viral Production Cells, and CTS LV-MAX Transfection Kit) that are all manufactured according to cGMP guidelines.
Do you have a GMP bank of the Viral Production Cells?

The Viral Production Cells are not cGMP-banked. However, we offer the CTS Viral Production Cells that are cGMP banked and optimized to work with the CTS LV-MAX Production Medium and CTS LV-MAX Transfection Kit.
Does the LV-MAX Production System Starter Kit work for Adeno-associated virus (AAV) production?

We have not optimized the LV-MAX Production System for AAV production. However, the Viral Production Cells contain the right genetic elements for producing AAV.

With the LV-MAX Lentiviral Production System, how do I scale up lentiviral production?

We have identified 3 key protocols that will address most customers' needs. In the user guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0017000_LV_MAX_ViralProductionSystem_UG.pdf ), you will find protocols for 1 mL in a 2 mL 96-well deep block, 5-30 mL in 50 mL conical tubes, and 30 mL-1 L in various shaker flask sizes. For larger-scale needs, we have developed a bioreactor protocol for up to 2 L. if you are interested, please email us at cts@thermofisher.com for details.
Do I need to optimize the LV-MAX Production System Starter Kit?

No, we've done the work for you. You will need to determine the best production cell density under your cell counting system. We recommend trying 3 different cell densities, i.e., 3 x 10E6, 4 x 10E6, and 4.5 x 10E6 cells/mL to determine the optimal cell density.

Does the LV-MAX Production System Starter Kit provide me with everything I need?

No. In addition to the LV-MAX System, you will need Opti-MEM I Reduced Serum Medium (Cat. No.31985088) as the DNA-LV-MAX Transfection Reagent complexation medium. You will also need a pLenti transfer plasmid and pLenti packaging mix. Please see the user guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0017000_LV_MAX_ViralProductionSystem_UG.pdf ) for a complete list of recommended materials and equipment.
For the LV-MAX Lentiviral Production System, I do not have the shaker recommended in the protocol, what are my options?

You may use an alternative shaker, but it must be CO2 resistant. If you use a non-digital shaker, you will need to optimize the shaker speed based on general observation. You must ensure that the cells do not settle but maintain suspended. You will need to generate your own cell growth curve to ensure you exceed 90% cell viability.

With the LV-MAX Lentiviral Production System, can I use a different lentiviral packaging system, that is not 3rd generation, instead of ViraPower Lentiviral Packaging Mix?

While we currently don't have data for prior generation lentiviral packaging systems, the LV-MAX Lentiviral Production System should be compatible with them but may require further optimization.
I am not achieving high titer using my own vector construct and the LV-MAX Lentiviral Production System. What factors influence my viral titer yield?

Several factors are known to affect viral yield:

- Production cell density: We suggest a cell density of 4 x 10E6 viable cells/mL in our protocol, but since different cell counting methods may give different counting results, we recommend that you optimize your production cell density based on your cell counting system. We recommend trying 3 different cell densities, i.e., 3 x 10E6, 4 x 10E6, and 4.5 x 10E6 cells/mL to determine the optimal cell density.
- Quality of the plasmid: Make sure that your plasmids are endotoxin-free and of high purity.
- Protocol: Following our cell culture protocol to establish your culture is the key to successfully use our LV-MAX System.
- Size of the gene: The size of the gene you are interested in will impact viral particle yield; the larger the size of the gene, the lower will be the viral particle yield.

Note: While viral titer yields are highly dependent on production volume, the specific gene being expressed, and its length, our R&D team as well as external collaborators have consistently seen an increase in viral titer that is equivalent to or 10-fold higher when using LV-MAX Lentiviral Production System as compared to adherent PEI, or suspension PEI methods.

How do I bank the Viral Production Cells?

Viral Production Cells can be frozen directly in cryogenic medium consisting of 90% LV-MAX Production Medium plus 10% DMSO. Here is the protocol:

- Bank cells at passage 3 when cell density reaches ˜3.0-4.5 x 10E6 viable cells/mL at >95% viability.
- Centrifuge the cells at 100 x g (low speed) for 5 mins to pellet. Discard the spent medium.
- Prepare freezing medium: e.g., 10 mL of freezing medium is 9 mL of LV-MAX Production Medium + 1 mL DMSO; mix well and sterilize by 0.2 µm filtration.
- Resuspend the cell pellet in the prepared freezing medium.
- Dilute the cells to a final density of 10 x10E6 viable cells/mL and aliquot 1 mL per cryovial.
- Freeze the cells at -80 degrees C for 1 day.
- Transfer frozen vials to liquid nitrogen for long-term storage.
Can I use other transfection reagents, such as PEI (polyethyleneimine), in the LV-MAX Lentiviral Production System?

LV-MAX Transfection Reagent, enhancer, and supplement are specifically designed to work synergistically with the LV-MAX Production Medium and Viral Production Cells to provide maximal lentiviral titer. Mixing and matching reagents is not recommended, as this will decrease the overall performance.

For Research Use Only. Not for use in diagnostic procedures.