Abstract
AGTC has developed a recombinant herpes simplex virus (rHSV-1)-based adeno-associated virus (AAV) production process. This method requires two rHSV-1 viruses; rHSV-1-rep/cap which expresses the AAV2 rep and cap proteins, and a second rHSV-1 that contains the therapeutic gene of interest flanked by AAV2 inverted terminal repeats (ITRs) necessary for rAAV production. Following co-infection of these two helper viruses onto HEK-293 cells, the rAAV DNA is replicated and packaged into rAAV capsids. rAAV is then liberated from the cells and purified. Our proprietary rAAV purification scheme removes all contaminants including HSV-1 proteins; however, it is necessary to develop a sensitive and quantitative method to confirm the absence of HSV-1 proteins within a level of detection, in the final rAAV preparation.
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