Adeno-associated Virus Production for Research

Cost-effective, scalable adeno-associated virus (AAV) vector production is important to meet commercial demand, and a smooth scale-up to clinical production is essential. To address these needs, we have created a complete AAV production system to help reduce production costs and streamline your transition from research to clinical scale.

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• High AAV titers—helps reduce production costs
• Scalability—suspension system with scalable protocols from shake flask to bioreactor scale
• Simplified workflow—streamlined protocol using helper virus-free triple transfection
• Animal origin–free (AOF)
• Clonal 293F-derived producer cells

Figure 1. Components of the Gibco AAV-MAX Helper Free AAV Production System.

The Gibco AAV-MAX Helper Free AAV Production System includes clonal HEK293F–derived suspension cells, chemically defined, animal origin-free, serum-free media, a proprietary transfection reagent, a booster, a novel production enhancer, and a polysorbate 20-based lysis buffer.

The AAV-MAX system provides a scalable and high-titer a platform for suspension AAV production. It is based on a high-density suspension culture of clonal 293F–derived Viral Production Cells 2.0 in Viral Production Medium. Optimal viral titers are mediated by our advanced lipid nanoparticle transfection reagent in combination with a novel production enhancer (Figure 1).

The components of the AAV-MAX system are designed to work synergistically, resulting in maximal titers and eliminating the need to optimize reagents and protocols. The complete system produces higher titers compared to swapping out individual components and using conventional polyethyleneimine (PEI)-based transfection reagents (Figure 2).

Figure 2. The complete system delivers maximum AAV titers compared to alternative transfection reagents. AAV2 and AAV6 were produced at a 30 mL scale in 125 mL shake flasks using the AAV-MAX system. These data highlight that the highest titers were achieved using the complete system compared to the reduced titers observed when using other transfection reagents. Titers were measured by qPCR. The data was normalized to the titer (vg/mL) of the complete AAV-MAX system.
 

The innovative AAV-MAX Helper-Free AAV Production System is a complete optimized suspension system that allows you to seamlessly and efficiently produce your AAV vector with high titers across multiple AAV serotypes.

Figure 3. The system yields high titers across multiple AAV serotypes. The AAV-MAX system was used to produce 5 AAV serotypes at a 30 mL scale in 125 mL shake flasks. These data show that high genome titers, high cell viabilities, and high viable cell densities (VCD) were achieved for each of the 5 serotypes. Titers were measured by qPCR.
 

The AAV-MAX system can help you obtain the same viral titer in less volume compared to PEI-based production systems, which translates to savings in media, downstream purification, lab space, and labor.

This advantage can mitigate contamination risks by working with smaller batch sizes. In addition, the AAV-MAX System requires less plasmid DNA per 1x10⁶ cells than alternative transfection reagents, which further drives production costs down.

Challenges associated with existing AAV production systems include low titers, high cost of cGMP plasmid DNA, low scalability, and lack of fit for purpose, cGMP reagents. The AAV-MAX system can help overcome these challenges by allowing you to produce high AAV titers using pre-optimized and regulatory-compliant* reagents that were developed for AAV production in a scalable suspension platform where volumetric viral titers are maintained as you scale up.

* Available with CTS AAV-MAX Helper Free Production System, set for release in 2022.

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