Application Note

Recombinant Adeno-Associated Virus Type 5 Production Process

Source: Cytiva

Human embryonic kidney 293 (HEK293) cells are susceptible to a broad range of viruses and can be easily transfected and are thus frequently used in the production of viral vectors and vaccines. Here, suspension adapted HEK293T cells and optimized triple plasmid transfection were used to produce rAAV, the main vector for gene therapy.

The key for a successful upstream production is having consistently high full capsid productivity, preferably using animal component-free suspension cell culture media. Important for the downstream process are efficient steps with good recoveries producing a high proportion of full capsids and low levels of impurities in the final bulk product. Scalable, cost-efficient, and robust upstream production, filtration, and chromatography-based processes are required for the purification of rAAV.

Depending on the clinical indication and administration route, the dose requirements and production volumes can vary significantly. Hence, the interest in scalable platforms for industrial rAAV production is increasing. Explore these results from a start-to-finish process for rAAV5 serotype, including virus production, harvest by cell lysis, clarification, concentration and buffer exchange, affinity capture, and finally anion exchange polishing to enrich or purify the full capsids.

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