Lentiviral vector production capabilities

Scalable solutions for lentiviral production

Whether you’re working with a suspension production system or an adherent production system, lentiviral production can be a costly part of your overall operation. Our goal is to help you maximize titer and ensure you have a cost-effective scalable solution for lentiviral vector production. Our lentiviral production methodologies utilize the trusted and widely cited family of Invitrogen Lipofectamine transfection reagents.

Suspension solutions for lentiviral production

The first optimized lentiviral production system for suspension culture

Gibco LV-MAX Lentiviral Production System
  • Cost-effective—reduce cost by more than 50%**
  • High titers—greater than 1 x 108 TU/mL (unconcentrated)
  • Research-grade and CTS options—seamlessly transition from discovery to commercial production

The Gibco LV-MAX Lentiviral Production System is the first optimized system that provides all the high-quality components you need to maximize your lentiviral production in suspension cultures in a chemically defined, serum-free environment. The Gibco LV-MAX Lentiviral Production System includes: HEK 293–derived suspension cells, serum-free media, proprietary transfection reagent, supplement, and our novel enhancer. Our HEK 293 derived viral production cells have been optimized for viral production in suspension culture.

One optimized and fully integrated system

Illustration of the seven stages from suspension culture to LV particles

The LV-MAX Lentiviral Production System key benefits include

Cost-effective and efficient lentiviral production

The LV-MAX system produced up to 15 times more virus than the PEI-mediated system in adherent cells and up to 10x more virus than in suspension cells (Figure 1), resulting in over 50% cost reduction compared to polyethylenimine (PEI)–based lentiviral production methods.

Current production method Switch to LV-MAX system and save*
Adherent PEI 58%
Suspension PEI 52%
* Cost comparison based on list price in USD and LV yield of 1 x 108 TU/mL using LV-MAX and 1 x 107 TU/mL using PEI-based adherent and suspension methods. Cost consideration includes media, transfection reagents, and culture vessels.
Bar chart demonstrating lentivirus titers are dramatically improved using LV-MAX Lentiviral production system

Figure 1. Lentivirus titers are dramatically improved using LV-MAX Lentiviral Production System. Unfiltered lentivirus produced by suspension cells using the LV-MAX Lentiviral Production System was compared with polyethylenimine (PEI)–mediated transfection of lentiviral vectors in adherent HEK 293T/FT cells and suspension HEK 293 cells. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells. The resulting cost savings is also shown.

Scale-up with confidence

The LV-MAX Lentiviral Production System is designed to support robust lentiviral production in a variety of suspension culture vessels. You can scale up or down based on your needs for greater throughput early in discovery or for seamless and efficient scale-up while maintaining high yield in a serum-free system (Figure 2). Regardless of scale, you can produce with more confidence.

bar chart demonstrating consistent lentiviral vector production from the LV-MAX system regardless of production format

Figure  2. LV vector was produced in different culture formats using the LV-MAX Lentiviral Production System. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells. Analysis of data showed no statistical significance between titers among the different production formats.

Multi-component system optimized for improved lentivirus production

The LV-MAX Lentiviral Production System provides a scalable and high-yield lentiviral vector production platform. It is based on a high-density suspension culture of HEK 293–derived viral production cells in LV-MAX Production Medium. Optimal viral titers are mediated by our advanced lipid nanoparticle transfection reagent in combination with a novel lentivirus-specific enhancer and supplement (Figure 3). All components work synergistically to help generate superior, functional lentiviral particles under xeno-free conditions compared to conventional polyethylenimine (PEI), serum-based culture systems (Figure 4). These data highlight the positive impact of using our complete system to maximize production of lentiviral vectors (LVV) compared to titers obtained when using incomplete or non-optimized systems (Figure 1).

Illustration of the 4 components of the LV-MAX lentiviral production system that result in 20x yields of lentiviral vector

Figure 3. LV-MAX Lentiviral Production System protocol overview.

Lentivirus production comparison for optimized kit versus PEI competitor

Figure 4. Significantly higher viral titers with LV-MAX system. Lentivirus was produced in 30 mL shaker flasks using our LV-MAX Lentiviral Production System or alternative transfection reagents and cells. LV293 = Gibco viral production cells (HEK 293–derived suspension cells), 293F = Gibco FreeStyle 293-F cell line, PEI = polyethylenimine.

ONE complete optimized system

In an effort to accelerate your research to clinical scale, we now offer a complete LV vector production system. The Gibco LV-MAX Lentiviral Production System produces up to 10x more lentivirus and is the first complete suspension production system.

graph of comparison of LV titers between research-grade and CTS (clinical) reagents
Figure 5. Comparison of LV titers between research-grade and CTS (clinical) reagents. LV vectors were produced in a 30 mL format in 125 mL shaker flask system. Performance of the LV-MAX Lentiviral Production System was shown to be equivalent to the CTS LV-MAX production system as measured by viral titers determined by transduction of HT1080 cells and flow cytometry analysis of GFP-positive cells.

Cost-effective and efficient lentiviral production

The LV-MAX system produced up to 15 times more virus than the PEI-mediated system in adherent cells and up to 10x more virus than in suspension cells (Figure 1), resulting in over 50% cost reduction compared to polyethylenimine (PEI)–based lentiviral production methods.

Current production method Switch to LV-MAX system and save*
Adherent PEI 58%
Suspension PEI 52%
* Cost comparison based on list price in USD and LV yield of 1 x 108 TU/mL using LV-MAX and 1 x 107 TU/mL using PEI-based adherent and suspension methods. Cost consideration includes media, transfection reagents, and culture vessels.
Bar chart demonstrating lentivirus titers are dramatically improved using LV-MAX Lentiviral production system

Figure 1. Lentivirus titers are dramatically improved using LV-MAX Lentiviral Production System. Unfiltered lentivirus produced by suspension cells using the LV-MAX Lentiviral Production System was compared with polyethylenimine (PEI)–mediated transfection of lentiviral vectors in adherent HEK 293T/FT cells and suspension HEK 293 cells. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells. The resulting cost savings is also shown.

Scale-up with confidence

The LV-MAX Lentiviral Production System is designed to support robust lentiviral production in a variety of suspension culture vessels. You can scale up or down based on your needs for greater throughput early in discovery or for seamless and efficient scale-up while maintaining high yield in a serum-free system (Figure 2). Regardless of scale, you can produce with more confidence.

bar chart demonstrating consistent lentiviral vector production from the LV-MAX system regardless of production format

Figure  2. LV vector was produced in different culture formats using the LV-MAX Lentiviral Production System. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells. Analysis of data showed no statistical significance between titers among the different production formats.

Multi-component system optimized for improved lentivirus production

The LV-MAX Lentiviral Production System provides a scalable and high-yield lentiviral vector production platform. It is based on a high-density suspension culture of HEK 293–derived viral production cells in LV-MAX Production Medium. Optimal viral titers are mediated by our advanced lipid nanoparticle transfection reagent in combination with a novel lentivirus-specific enhancer and supplement (Figure 3). All components work synergistically to help generate superior, functional lentiviral particles under xeno-free conditions compared to conventional polyethylenimine (PEI), serum-based culture systems (Figure 4). These data highlight the positive impact of using our complete system to maximize production of lentiviral vectors (LVV) compared to titers obtained when using incomplete or non-optimized systems (Figure 1).

Illustration of the 4 components of the LV-MAX lentiviral production system that result in 20x yields of lentiviral vector

Figure 3. LV-MAX Lentiviral Production System protocol overview.

Lentivirus production comparison for optimized kit versus PEI competitor

Figure 4. Significantly higher viral titers with LV-MAX system. Lentivirus was produced in 30 mL shaker flasks using our LV-MAX Lentiviral Production System or alternative transfection reagents and cells. LV293 = Gibco viral production cells (HEK 293–derived suspension cells), 293F = Gibco FreeStyle 293-F cell line, PEI = polyethylenimine.

ONE complete optimized system

In an effort to accelerate your research to clinical scale, we now offer a complete LV vector production system. The Gibco LV-MAX Lentiviral Production System produces up to 10x more lentivirus and is the first complete suspension production system.

graph of comparison of LV titers between research-grade and CTS (clinical) reagents
Figure 5. Comparison of LV titers between research-grade and CTS (clinical) reagents. LV vectors were produced in a 30 mL format in 125 mL shaker flask system. Performance of the LV-MAX Lentiviral Production System was shown to be equivalent to the CTS LV-MAX production system as measured by viral titers determined by transduction of HT1080 cells and flow cytometry analysis of GFP-positive cells.

Our commitment to quality

Our products are backed by our global scale, our regulatory and technical support teams, an unrelenting focus on quality, and decades of experience with good manufacturing practices (GMP). This winning combination has enabled us to become a chosen partner of many top 50 biopharmaceutical companies worldwide. All of our manufacturing sites have independently been certified by the Quality Management Systems (ISO 9001 and/or ISO 13485). Gibco Cell Therapy Systems (CTS) LV-MAX Lentiviral Production System is manufactured in conformity with GMP for medical devices (21 CFR Part 820), and follows USP <1043>* and European Pharmacopoeia (Ph. Eur.) 5.2.12 recommendations. All CTS products come with a Drug Master File in the USA/CAN/JP or a Regulatory Support File in other regions.

We are dedicated to your commercial success and that is why we are continuing to develop new lentiviral production solutions and invest in our GMP manufacturing. Our commitment to quality systems and materials, redundant manufacturing, and continuity of supply is upheld through our supply agreements, risk assessment, and mitigation strategies. Utilizing these practices enables our customers to be successful, helping clear the path to commercialization. Our mission is to help you meet current and future viral vector demand and clear the path to commercialization to help you stay at the forefront of cell and gene therapy. 

Tell us how we can help offer you the best solution for your lentiviral vector production needs

ONE suspension lentiviral production system from research to commercial manufacturing

A smooth ramp-up from research to clinical production is essential. In an effort to accelerate your development timelines, we offer both research-grade and GMP solutions for ONE seamless transition.

Attributes LV-MAX CTS LV-MAX
Intended use For research use only For research use or manufacturing of cell, gene, or tissue-based products
Pack sizes Starter kit (300 mL) >10 L
Formulation Same Same
Performance Same Same
Packaging cell line Research use GMP-banked HEK 293
Extensive cell line documentation
Regulatory support documentation None DMF
Regulatory Support File
GMP quality Manufactured to ISO 9001 or ISO 13485 Manufactured to ISO 13485 21 CFR Part 820/USP <1043>

NEW GMP banked HEK 293 cells for viral vector production

GMP banked HEK293 cells for viral vector production

CTS Viral Production Cells are derived from the human embryonic kidney (HEK) 293 cell line and were adapted to suspension culture in a chemically defined medium. These cells were developed to support cell and gene therapy applications, particularly intended for viral vector manufacturing. To comply with recent FDA CMC guidance for Human Gene Therapy INDs, CTS Viral Production Cells do not contain SV40 large T antigen nor have they been genetically engineered. With detailed cell line lineage documentation and extensive quality control testing, you can seamlessly transfer these GMP cells to clinical production. These cells are optimized to culture in suspension with the CTS LV-MAX Production Medium. For more information on this cell line, please contact outlicensing@thermofisher.com.

  • Fully documented cGMP-banked cell line (HEK 293F derived) with detailed cell line lineage history 
  • Absence of SV40 large T antigen
  • Supports >10 million cells/mL in chemically-defined medium

Ordering information for LV-MAX kits and components


Advanced lipid nanoparticle technology for superior lentiviral production in adherent cultures
Invitrogen Lipofectamine 3000 Transfection Reagent is a highly efficient, cost-effective tool for lentiviral production. This versatile reagent enables high viral titers even with genes that are large or difficult to package.

Invitrogen Lipofectamine 3000 Reagent

  • High titers—including genes that are large or difficult to package
  • Gentle—reduced reagent dose for improved cell viability
  • Flexible—compatible with your existing protocol
  • Cost-effective—use less ancillary components and labor
Invitrogen Lipofectamine 3000 reagent

In a side-by-side comparison between Lipofectamine 3000 Reagent, Lipofectamine 2000 Reagent and PEI, Lipofectamine 3000 reagent–mediated lentiviral production was ~5–10 times greater than that obtained from the other transfection reagents (Figure 5).

bar chart showing lentivirus production in adherent cells with Lipofectamine 3000 reagent

Figure 5. Lentivirus production in adherent cells with Lipofectamine 3000 reagent. pLenti6.3/V5-GW/EmGFP and Invitrogen ViraPower Lentiviral Packaging Mix were delivered by Lipofectamine 3000 reagent, Lipofectamine 2000 reagent, and polyethylenimine (PEI) in a 6-well culture plate with HEK 293T/FT cells. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells.

Is my CTS LV-MAX transfection reagent supposed to be cloudy?
It is normal to see some turbidity and cloudiness in LV-MAX transfection reagent. Lipid transfection reagents are sensitive to low temperature; if you store them at temperatures lower than the recommended 2–8° C storage conditions or if the reagent freezes it will precipitate, leading to low transfection efficiency or inactivity.

Is my LV-MAX transfection reagent missing a plastic seal around its lid?
The LV-MAX transfection reagent does not come with an external plastic cap seal. This avoids exposing the final product to temperature fluctuations experienced during the seal application process. This does not indicate a breach in container closure integrity.

Do I need to obtain licensing rights if I just want to try the system?
Please refer to our standard sales terms and conditions and limited use label license for specific products. For additional information, please contact us at outlicensing@thermofisher.com.

Do you have GMP-banked cells that can be used for clinical work and IND filing?
Yes, we offer cGMP-banked viral production cells that have been characterized according to ICH Q5a guidance.

What is the intended claim for these products?
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 individual product for further details on the intended use claim.

What comes with the CTS Viral Production Cells?
We have extensive documentation to demonstrate lineage history, viral clearance, and traceability of our GMP manufactured process. Under CDA, we can share our Certificate of Analysis and Table of Contents. Full access to documentation package is available through outlicensing@thermofisher.com.

Can I use the CTS Viral Production Cells or CTS LV-MAX system for clinical trials or commercial use?
Please refer to our standard sales terms and conditions and limited use label license for specific products. For further inquiries, please contact us at outlicensing@thermofisher.com.

What are the major differences between LV-MAX and the CTS LV-MAX products?
Performance between the two systems is equivalent. 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 complimented 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.

Are the cells clonally expanded?
Viral Production Cells were derived from Freestyle 293-F cells (Cat. No. R79007), which wasn’t clonally generated. The lineage of our current Viral Production Cells at one point was clonally derived. For further details on our cell line lineage, please contact outlicensing@thermofisher.com.

Do you have any guidelines for viral purification under serum free conditions?
For downstream purification of virus, we recommend Tangential Flow Filtration. We do not recommend ultracentrifuge for serum-free conditions to maintain integrity of viral vectors. Please contact cellculturesupport@thermofisher.com for suggested protocol.

Do you have a protocol to detect residual components of the LV-MAX transfection reagent and enhancer?
We can provide protocol guidance for how to detect transfection reagent components under CDA. Please contact cellculturesupport@thermofisher.com for information.

I'm interested only in the transfection reagent. Can I purchase it separately from the transfection kit?
Not at this time. The LV-MAX Transfection Kit comes with three components that are not sold a la carte. The system components were specifically optimized to work as a system for maximal viral vector titer. We can however customize different ratios of the supplement, enhancer and transfection reagent.

LV-MAX is more expensive than PEI-based transfection reagent. How can you claim it’s a low cost option?
The cost per viral particle is much lower using LV-MAX system compared to PEI-based method due to the robustness of the LV-MAX system. Because of the high efficiency of the complete system, less material and volumes are needed to make the same amount of virus, which also reduces labor and ancillary consumable reagents (plasmids, purification etc.).

Have you tried other media like HEK 293 expression media with your system?
Yes, we have tried other media and have seen reduced performance. For maximal LV vector yield, we recommend using LV-MAX medium as part of the entire LV-MAX system.

Have you tried other commercially available transfection reagents (i.e., PEI) in your system?
Yes, we have tried a full range of transfection reagent formulations while developing the system. LV-MAX Transfection Reagent is compatible with the suspension Viral Production Cells to support transfection in a high cell density format and offers higher performance than other transfection reagents.

My cells are initially low viability post thaw, is this normal?
The protocol recommends culturing the cells to passage 5 (P5) so that the cells recover sufficiently prior to starting the protocol for LV production. It is normal to see variance in viability upon thaw, but this should normalize during recovery passages.

What scale have you tested for scale up so far? Do you have protocols you can share?
We continually develop large scale protocols to support clinical and commercial LV vector production. Please contact cellculturesupport@thermofisher.com for the latest information.

What density should I be seeding the cells?
Depending on whether you are doing a 3 day or 4 day culture, we have outlined the differences in seeding densities in our LV-MAX user guide manual. We also have specific guidance around seeding densities to be used for lentiviral vector production.

Why do we have to dilute the cells once more before transfection? Is that necessary?
This is a necessary step to replenish nutrients to the cells and ensure that the cells are at the correct density to achieve the concentrations needed for optimal transfection efficiency. Proper cell density at time of transfection ensures maximal performance of the system.

Can I transfect before passage 5 (P5)?
We recommend that cells be used for lentiviral production following passage 5 to allow for full recovery of the cells from the cryopreservation.

The Viral Production Cells that come with the starter LV-MAX Lentiviral Production System are supposed to be good in passage 5–20. Does this refer to passages 5–20 post-thaw, or should we consider the number of passage the cells went through before cryopreservation??
The passages should be counted upon thaw, and discarded when they reach passage 20 post-thaw.

Should I harvest at 24 hours and 48 hours? What about waiting for 72 hours? What’s the best harvest method?
Our system has been optimized to provide maximal lentiviral production at 48 hours post transfection. Waiting until 72 hours is not recommended since existing particles within the media will infect the production cells and can reduce the titer.

What is the recommended ratio of plasmids to use in the system?
We recommend using the lentiviral packaging plasmids to the transfer plasmids at a ratio of 3:2.

Do you have recommendations for how to measure non-fluorescent LV titer?
In our user guide manual, we provide a protocol for titer measurement using antibiotic selection.

Why is there no media exchange requirement in the LV-MAX protocol and the cells remain in the same media throughout the two-day transfection? What data do you have to check on the effect of media exchange??
We have done comprehensive studies on the media changes. The LV-MAX production medium is specially designed to support the health of the Viral Production Cells cultured at high-density. Therefore, there’s no need for media exchanges.

Enhancer addition time is suggested to be anytime between 5–14 hours after transfection, how critical is it that we stay within this time frame window?
Enhancer addition was tested over the entire rage of 5–14 hours and was found to perform with equivalent efficiency when added anytime during that range of time. The enhancer formulation coupled with the broad window of addition time was specifically optimized to allow the user flexibility in schedule and start time of the assay without affecting performance.

Have you tried other plasmids with your systems?
The LV-MAX production system is designed to be compatible with any format of plasmid DNA. The quality of plasmids (e.g., purity), ratio of the plasmids, and vector design all contribute to viral titer. Please contact us at cellculturesupport@thermofisher.com for suggestions.

I need support in designing my plasmid constructs. Who should I reach out to?
Please contact our GeneArt department for assistance at support@geneart.zendesk.com.

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  • Lentiviral Gene Delivery for Mammalian Expression Support
    Explore our resources, tip and tricks, and troubleshooting advice for delivering your lentiviral construct into mammalian cells for expression of your protein of interest. Obtain detailed information on every step of the workflow, including preparation of the lentiviral construct, generation of the lentiviral stock, viral titering, transduction, and analysis of expression.

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*CTS products are manufactured to meet the ancillary material supplier responsibilities for cell-, gene-, and tissue-engineered products. Other aspects of USP <1043> are the responsibility of the end user to assess.
** Compared with PEI-based lentiviral production methods.

Intended use of the products mentioned on this page vary. For specific intended use statements please refer to the product label.