Production of recombinant adeno-associated virus 5 using a novel self-attenuating adenovirus production platform

Recombinant adeno-associated virus (rAAV) has become a prominent vector for clinical use. Despite an increase in successful clinical outcomes, the amount of high-quality rAAVs required for clinical trials and eventual commercial demand is difficult to produce, especially for genetic diseases that ar...

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Bibliographic Details
Published in:Molecular therapy. Methods & clinical development 2024-09, Vol.32 (3), p.101320, Article 101320
Main Authors: Roach, Matthew K., Wirz, Phillip, Rouse, Jeremy, Schorzman, Allison, Beard, Clayton W., Scott, David
Format: Article
Language:English
Subjects:
AAV
adeno-associated virus
adenovirus
potency
production
purification
TESSA
triple transfection
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Summary:Recombinant adeno-associated virus (rAAV) has become a prominent vector for clinical use. Despite an increase in successful clinical outcomes, the amount of high-quality rAAVs required for clinical trials and eventual commercial demand is difficult to produce, especially for genetic diseases that are prevalent or require high doses. Many groups are focused on establishing production processes that can produce sufficient rAAV while maintaining potency and quality. Our group used a novel production platform to increase our yield of rAAV5. This production platform uses tetracycline-enabled self-silencing adenovirus (TESSA) to deliver the wild-type AAV replication and capsid genes alongside the adenovirus helper genes necessary for production. Here, we describe our efforts to evaluate the TESSA platform in house. We conducted numerous experiments to determine the optimal conditions for producing rAAV5 from the TESSA production system. We then produced rAAV5 from the TESSA system to compare against rAAV5 produced from triple transfection. Ultimately, we generated data that showed that the vector genome yield of rAAV5 produced with TESSA was >20-fold higher than rAAV5 produced with triple transfection. Additionally, our data show that quality as well as potency in mice of rAAV5 produced with the TESSA system and by triple transfection are equivalent. [Display omitted] Roach and colleagues evaluate a novel modified adenovirus production platform to produce rAAVs. The authors demonstrate TESSA produced AAV yields >20-fold that of triple transfection-produced AAVs.
ISSN:2329-0501
2329-0501
DOI:10.1016/j.omtm.2024.101320