Generation of clinical grade recombinant adeno-associated virus

• Main Author: Tie, Mark
• Other Authors: Offner, Gwynneth
• Language: en_US
• Published: 2018
• Subjects: Biology
• Online Access: https://hdl.handle.net/2144/31279

Recombinant adeno-associated viruses (rAAV) remain one of the most encouraging gene therapy vectors for treating patients with genetic abnormalities. rAAV can safely deliver long-lasting expression of a therapeutic transgene to a wide range of cell types. One challenge with therapeutic rAAV is the ability to generate enough virus for clinical trials and commercial supply. Doses administered systemically for neuromuscular or blood disorders can exceed 1 x 1014 viral genomes per patient. Approximate yields from a rAAV production are around 1 x 104 viral genomes per cell, meaning batch cell numbers would need to exceed 1 x 1010 for a single dose. This amount of therapeutic virus will require a production platform that can reliably generate sufficient quantities of therapeutic rAAV to meet patient demand. Common expression platforms used in academia and industry are insufficient for generating this amount of virus, let alone in a controllable or reproducible setting. More advanced systems based on stable cell lines paired with alternate viruses offer avenues for achieving more efficient production of virus, however there is no clear agreement on which system is most amenable to regulatory approval while also delivering safe and cost efficacious therapies to patients. This paper will outline the basic biology of AAV and rAAV vectors. The common methods used to produce rAAV will be evaluated and a roadmap for producing clinical grade rAAV at scale will be provided.