Optimisation of retroviral production systems for gene therapy applications

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http://eprints.org/relation/isVersionOf http://etheses.bham.ac.uk/id/document/21712 http://eprints.org/relation/isVolatileVersionOf http://etheses.bham.ac.uk/id/document/21712 http://eprints.org/relation/isIndexCodesVersionOf http://etheses.bham.ac.uk/id/document/21712 archive 4031 disk0/00/00/35/92 2012-07-13 11:20:31 2019-01-23 09:56:37 2012-07-13 11:20:31 thesis show 0 Warnock James Neill Al-Rubeai Mohamed University of Birmingham Optimisation of retroviral production systems for gene therapy applications unpub RM TP 30sch_engi Retroviral vectors are a promising tool for gene therapy. However, there are two major problems to overcome if a viable commercial production process is to be established. These are the instability of virus particles and the low virus titres. The characteristics of the producer cells were determined in batch culture, semicontinuous culture and semi-continuous culture at 32°C. Additionally, cell attachment, growth and virus production on various macroporous microcarriers was assessed under static and stirred conditions. Alternative strategies for the cultivation of cells were also investigated. These included spinner basket, packed bed and spinner flask cultures with semi-continuous feeding and packed bed, fixed bed, fluidised bed and stirred tank cultures with continuous perfusion of culture medium. Of these the fixed bed bioreactor had the highest cell specific productivity and was capable of running for 28 days. The fluidised bed bioreactor had the highest reactor productivity, due to the higher cell number. Optimisation of culture medium was performed with regard to serum concentration. The greatest production was observed at an initial serum concentration of 2.5% (v/v). The findings in this thesis will assist the development of an efficient method for the production of clinical grade retro viral vectors for gene therapy applications. 2003-07-22 2002-11 graduation 2261484 eng other Scanned by Hollingworth and Moss This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder. public 239 University of Birmingham School of Chemical Engineering d_ph na 2003-07-22 graduation