Retroviral vector production for gene therapy applications

[摘要] The production of retroviral vectors for gene therapy applications faces a number of challenges. Of primary concern is the low titre of vector stocks produced by packaging cells in culture and the inherent instability of retroviral vector activity. A systematic investigation of culture parameters that can effect vector titre was conducted. Physical and chemical factors including temperature, pH, medium composition, dissolved oxygen and serum concentration were all assessed. In addition, a number of studies were undertaken to assess the effects of packaging cell growth rate on vector production. The use of a packed bed, as a novel system for large-scale vector production, was also investigated. Prolonged production of retroviral vector stocks was demonstrated in the packed bed system with immobilised packaging cells. Determination of the critical culture parameters allowed optimisation of culture conditions, which can be continuously controlled in the packed bed system, thereby ensuring optimal vector production throughout the production period. Furthermore, vector decay rates can be reduced by the immediate collection of vectors into a recovery vessel. The studies within this thesis will aid the development of appropriate procedures for the large-scale production and handling of retroviral vector stocks for human gene therapy applications.

[发布机构] University:University of Birmingham;Department:School of Chemical Engineering

[关键词] Q Science;QH Natural history;QH426 Genetics