The current standard for manufacturing GMP-grade Adeo associated viral (AAV) vectors rely on a tri-plasmid system that involves transient transfection of mammalian cells with three plasmids carrying the necessary packaging components. While this method is versatile and relatively fast, scaling it for clinical applications is challenging. The need to transfect all three plasmids into a single cell, the high quantity of plasmid DNA (#pDNA) required, and the necessity of ratio optimization among the plasmids introduce variability, limit efficiency, and increase time and labor costs.

💡To address these issues, we developed an all-in-one single-plasmid system, #AAVone, which consolidates all necessary elements—adenovirus helper genes (E2A, E4orf6, VA RNA)AAV helper genes (rep, cap), and the AAV vector genome—into a single compact plasmid with a 13-kb backbone plus the AAV genome. By simply transfecting the #pAAVone plasmid into host cells, high yields of AAV vectors can be produced, achieving
🔹 Over 1×10^15 vector genomes per liter (vg/L) in suspension cells,
🔹 Representing a 2- to 4-fold increase in yield compared to the tri-plasmid system.

💡AAV vectors produced using #AAVone exhibit similar capsid composition, genomic pattern, ITR integrity, and infectivity in mouse model, with fewer or comparable plasmid- and host cell-related impurities. Replication-competent AAV (rcAAV) testing showed no detectable rcAAV in 1×10^9 vector genomes.

💡#AAVone is versatile across various cell types and transfection reagents, and it demonstrates low batch-to-batch variation with easy scalability, Moreover, it requires fewer plasmid DNA and produces higher full-particle ratios, making it an efficient and cost effective solution for GMP-grade AAV vector production.

💁‍♀Benefits of using #AAVone system, it simplifies #AAV #packaging by using a single plasmid and omitting the plasmid ratio optimization step;
🔹 Easy scalable from flasks to bioreactors;
🔹 Improves AAV qualities evidenced by increased full to empty AAV ratio, decreased plasmid- and host genome-related impurities, ITR and AAV genomic integrity;
🔹 Requires 50%-75% less input plasmid use and up to 4 times more in AAV production with improved qualities;
🔹 Effectively produce safe AAV vectors with little to no rick of forming rcAAV
🔹 Generated vectors show equal to better performance in mouse models compared to that of tri-plasmid.

🌐 Learn more about the groundbreaking AAVone system at: https://sabrinahe22de09a181.wpcomstaging.com

#AAVone #GeneTherapy #AAV #Biotech #Innovation #GMP #ViralVector #BiotechSolutions