Traditional methods for the production of adeno-associated viral (AAV) vectors — crucial components in gene therapies — rely on plasmid DNA (pDNA), which can pose safety concerns due to bacterial sequences that might trigger immune responses and are fraught with inefficiencies that escalate production costs and timelines. Touchlight’s enzymatically synthesised dbDNA addresses these safety and quality concerns by completely eliminating bacterial sequences from the DNA used, thereby eliminating the need for antibiotic resistance assays. This not only enhances the safety profile by reducing the risk of unwanted immune reactions but also improves the overall particle quality and integrity of viral vectors. Additionally, the reduction in required DNA and transfection agents in the dbDNA method enhances yield and further lowers production costs, making it a safer and more efficient alternative for gene therapy vector production.

• Transfection Reagent & DNA Savings: dbDNA operations use up to 50% less transfection reagent compared to traditional methods.
• Expedited Production Time: Faster production timelines, with GMP-grade material deliverable within 5-6 weeks.
• Broad Applicability: dbDNA can carry sequences up to and over 30 kilobases, making it valuable for AAV vector, mRNA therapy, gene-editing, and DNA vaccine applications.

This innovation not only accelerates the initial evaluations of AAV vectors with readily available research-grade dbDNA but also offers GMP-grade dbDNA on expedited timelines, paving the way for faster and more cost-effective development of AAV-based therapeutics. Furthermore, the scalability of the dbDNA platform allows for significant production volumes more cost-effectively, streamlining the AAV vector production process, and thereby accelerating the progress in gene therapy from preclinical stages to commercial scale.

Traditional bacterial fermentation techniques used to manufacture plasmid are time-consuming, expensive, and inherently unstable. The production of sufficient GMP grade material thus imposes a major bottleneck on industrial-scale manufacturing of lentiviral vectors (LVV). Touchlight’s linear doggybone DNA (dbDNA) is an enzymatically amplified DNA vector produced with exceptional speed through an in vitro dual enzyme process, enabling industrial-scale manufacturing of GMP material in a fraction of the time required for plasmid.

This latest Nature Gene Therapy paper demonstrates that dbDNA could be optimised for the manufacture of high titre LVV and that dbDNA technology can provide a highly effective solution to the plasmid bottleneck.