Sponsored content brought to you by
Gene therapies offer incredible promise for treating diseases with a genetic component. Several gene therapies are already available, and adeno-associated virus (AAV) has become a popular choice for delivering genes. Thus far, scientists have produced approved therapies using triple-plasmid transient transfection and adherent cells. But many envision a future where producer cell lines will take over for large-scale production. Here’s what to know about AAV producer cell lines for gene therapies.
Definitions vary
Producer cell lines are genetically engineered to stably express the genetic elements required to produce the AAV of interest in small and large-scale manufacturing platforms. A true AAV producer cell line doesn’t require helper virus infection or plasmid transfection.
Developing a stable AAV producer cell line takes skill
A stable AAV producer cell line incorporates four elements required for AAV production — Rep, Helper, capsid, and the gene of interest — into the host genome. These cells must be skillfully engineered to support robust growth, viral titer, and quality. At Cytiva, we have developed a clonal HEK293 alpha cell line and a clonal CAP™ alpha cell line derived from human amniocytes.
Both have Rep and Helper functions stably integrated into their genomes and are used as the basis for generating producer cell lines. We do single-cell cloning from the producer pool and use high-throughput screening to select the best producer clones for productivity, packaging efficiency, and cell culture profile.
AAV producer cell lines offer process benefits
First, stable AAV producer cell lines simplify production by eliminating the need for transient transfection and helper virus infection. Next, such a cell line helps to minimize batch-to-batch variation because all genes are stably incorporated into the genome, leading to each cell producing viral vectors in the same manner. Stable producer cell lines also support manufacturing scales in the thousands of liters, whereas transient transfection at such scales is costly and technically challenging. Finally, using them removes the need to source large quantities of GMP-grade plasmids and transfection reagent — good news for supply chain predictability.
They support lower cost of goods manufactured
The cost of plasmids and transfection reagent comprises roughly 40% of the total cost to manufacture AAV without a producer cell line. Eliminating these expensive components has the potential to lower the costs of making a batch.
Learn more about AAV and our gene therapy workflow solutions at cytiva.com.
