Adeno-associated virus (AAV) have great potential as delivery vehicles for gene therapy for its many benefits: long-term transgene expression, high stability, transduction of large variety of cells, lack of pathogenicity and low immunogenicity.
Various strategies are developed for AAV vector production, one of the most common being the transient transfection of several plasmids into mammalian cells. This strategy is complex by the number steps required to recover the final product: plasmid production and purification, cell expansion, plasmid transfection, AAV production and purification.
Many challenges need to be addressed for the use of AAV in human gene therapy. Robust and scalable bioproduction process is required to yield high quantities of AAV vector to answer clinical development needs. Consistent viral particles production as well as proper characterization of AAV vector is essential to ensure safety and efficacy of the end-product.
Here at FHNW, we produce and purify plasmids required for AAV production with our in-house developed plasmid platform. We evaluate different conditions for triple plasmid transfection to increase AAV yield in suspension and set up robust and reliable purification methods to later characterize the produced AAV particles.
Motivated students have the chance to acquire essential skills in both upstream and downstream processing (USP and DSP) during internships such as: microbial and mammalian cell expansion, bioreactor operation at small-scale (2L) up to larger scales (20L), plasmid transfection, viral vector production, setup of chromatographic techniques, Design of Experiments (DoE), data analysis using Python, etc. These techniques prepare students well for a smooth transition into various positions in the biopharmaceutical industry.