Whether you’re working with an adherent or a suspension 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.
Our products are backed by our scale, ISO 13485- and ISO 9001-certified manufacturing sites, decades of experience with good manufacturing practices (GMP) and our regulatory and technical support teams. We are dedicated to your commercial success and that is why we are continuing to develop new lentivirus production solutions and invest in our GMP manufacturing. 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.
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.
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*
* 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.
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 HEK293 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.
Figure 2. Scalability of LV-MAX Lentiviral Production System in HEK 293–derived suspension cells. Lentiviruses were produced in different volumes ranging from a 96 deep-well plate to a 2 L bioreactor using LV-MAX Lentiviral Production System. The lentiviral titer was determined by transducing HT1080 cells and analyzing GFP-positive cells; similar titers were observed in different volume 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).
Figure 3. LV-MAX Lentiviral Production System protocol overview.
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.
GMP banked HEK293 cells for viral vector production
CTS Viral Production Cells are derived from the human 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 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 LV-MAX Production Medium. For more information on this cell line, please contact email@example.com
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
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).
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.
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.