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The Genesis of Gendicine: The Story Behind the First Gene Therapy

therapeutics using an adenoviral vector.

I used the Cell-Cube (parallel-plate reactor) system in the lab, but yields were too low, and we had cell growth problems. We produced approximately 1 x 1014 VP in a medium sized Cell-Cube (21,250 cm 2 surface area). When we installed a 14-L CelliGen Plus packed-bed bioreactor (from New Brunswick Scientific) in our manufacturing facility, we produced 15 times more than in the Cell-Cube system. Approximately 2 x 1015 VP can be produced in a 14-L bioreactor that is partially loaded with 200-g Fibra-Cel disks, and 1 x 1015 VP adenoviral vector in a 5-L bioreactor loaded with 100-g disks. We observed two periods of peak virus release into the supernatant on day 3 and day 5 postinfection, respectively. The viral bursts of 40,000 to 50,000 viral particles/cell were attained on day 3 postinfection.

A complete panel of lot-release qualitycontrol criteria for the final product has been established. Those criteria include vector purity, particle concentration, infectivity, gene expression, potency, and product safety criteria such as sterility, adventitious viruses, and level of replication-competent adenovirus (RCA). For example, our released final product has the following quality characteristics: IU/VP ratio of about 4.8%, purity over 97%, and less than 1 RCA in 3 x 1010 VP.

BPI: What is the source of cells?

Peng: Gendicine is based on an E1-deleted adenoviral vector containing the p53 tumorsuppressor gene. Engineered cells expressing adenovirus E1 proteins are required to produce Gendicine. Usually, people use the HEK 293 cell line for this purpose. We have subcloned a cell line called SBN-Cel from the 293 cell line. SBN-Cel is much better tha
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