Preventing spontaneous genetic rearrangements in the transgene cassettes of adenovirus vectors.

摘要: First-generation, E1/E3-deleted adenoviral vectors with diverse transgenes are produced routinely in laboratories worldwide for development of novel prophylactics and therapies a variety applications, including candidate vaccines against important infectious diseases, such as HIV/AIDS, tuberculosis, malaria. Here, we show, two different (both encoding malarial antigens) inserted at the E1 locus, that rare viruses containing transgene-inactivating mutation exhibit selective growth advantage during propagation E1-complementing HEK293 cells, they rapidly become major or sole species viral population. For one these transgenes, demonstrate yield cytopathic effect enhanced by repression transgene expression producer cell line, using tetracycline repressor system. In addition to mutations, which occurred pre-viral genomic clone bacteria, other after reconstitution describe types mutation, small deletion gross rearranging duplication, studied. These were uncertain origin, effects on not fully characterized. We that, together minor protocol modifications, cells enables production genetically stable chimpanzee adenovirus vector expressing antigen had previously been impossible derive. results have implications basic pre-clinical studies derivation products destined large-scale amplification biomanufacture.