Quick, Cost-Effective Approach for Manufacturing Multiple Vectors for Gene Therapy and Biological Therapeutics
This production of AAV enables two, or potentially more, vectors to be produced, characterized, and delivered as a single clinical therapeutic product and could be used as a service for manufacturing large amounts of viral vectors. AAV vectors are typically created using helper plasmids to package a plasmid comprising of a gene of interest within a capsid, or a protein shell that encloses the genetic material. However, each capsid can only hold a single copy of gene, so if two vectors are required they must be produced separately and then delivered separately, resulting in a loss of time and an increase in the production cost and time. Many diseases that are candidates for gene therapy treatment require two separate AAV vectors to be produced and delivered since the gene of interest is often too large for packaging in AAV. University of Florida researchers have developed a single step production procedure that allows for the production of multiple vectors simultaneously, thus forming a single therapeutic clinical product. This improvement not only creates a single product and provides an approach for packaging vectors, but it also greatly reduces the production output necessary to manufacture AAV vectors and saves production time.
Single lot production of AAV that packages multiple vectors, which can be characterized and administered as a single product
- Reduces the production output necessary to manufacture viable amounts of virus, saving production time
- Decreases time necessary to acquire the vectors, allowing AAV vectors to be administered considerably faster
- Allows for agencies to evaluate two vectors as a single product, significantly reducing the amount of oversight and redundant testing of multiple vectors
This AAV production approach offers an improvement in the manufacturing of large amounts of viral vectors, because it allows multiple vectors to be created, classified, and administered as one single product. AAV vectors are produced using a plasmid containing a gene of interest as well as two helper plasmids that supply an adenovirus helper gene and the capsid, which encloses the genes in a protein shell. Due to the nature of vector packaging, each capsid can contain only a single copy of the gene of interest. Researchers at the University of Florida have developed a way to exploit this fact by packaging the gene of interest in equivalent stoichiometric ratios depending on the input concentrations of plasmids. In other words, the amount of full-capsid vector produced is directly related to the input ratios of transgenes, capsids, and helper plasmids. By combining plasmids at known concentrations while maintaining the ability for independent characterization, AAV vectors can be manufactured and delivered as a single clinical therapeutic product.