Modeling Retrovirus Production for Gene Therapy. 1. Determination of Optimal Bioreaction Mode and Harvest Strategy

Although retroviruses are a promising tool for gene therapy, there are two major problems limiting the establishment of viable industrial processes: retrovirus stability and low final yield in the supernatant. This fact emphasizes the need for an effective process optimization, not only at a genetic...

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Bibliographic Details
Published in:	Biotechnology progress Vol. 16; no. 2; pp. 213 - 221
Main Authors:	Cruz, Pedro E., Almeida, Jonas S., Murphy, Patrick N., Moreira, José L., Carrondo, Manuel J. T.
Format:	Journal Article
Language:	English
Published:	USA American Chemical Society 2000 American Institute of Chemical Engineers
Subjects:	Additives Animals Batch cell culture Biological and medical sciences Bioreactors Biosynthesis Biotechnology Biotechnology - methods Butyrates - pharmacology Cell Culture Techniques - methods Cell Division - drug effects Cells, Cultured - virology Dexamethasone - pharmacology Fundamental and applied biological sciences. Psychology Gene therapy Genetic Therapy - methods Half-Life Health. Pharmaceutical industry Industrial applications and implications. Economical aspects Mathematical models Models, Biological Moloney murine leukemia virus - genetics Moloney murine leukemia virus - growth & development Optimization Patient treatment Reproducibility of Results Retroviridae - genetics Retroviridae - growth & development Retrovirus Sensitivity and Specificity Temperature Viruses Cell culture Recombinant virus Rodentia Downstream processing Retroviridae Modeling Optimization Stirred tank reactor Virus Vertebrata Bioreactor Cell line Mammalia Batchwise Mouse Perfusion Animal Production Mathematical model Gene therapy Application Fibroblast Vector
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Summary:	Although retroviruses are a promising tool for gene therapy, there are two major problems limiting the establishment of viable industrial processes: retrovirus stability and low final yield in the supernatant. This fact emphasizes the need for an effective process optimization, not only at a genetic level but also at a bioprocess engineering level. In part 1 of this paper a mathematical model was developed to optimize the bioreaction yield by determining the best retrovirus harvest strategy in perfusion cultures. PA317 cells producing recombinant retroviruses were used to develop and test this model. Cell culture was performed in stirred tanks using porous supports. The parameters of the proposed model were experimentally determined for batch and perfusion cultures at 32 and 37 °C both with and without additives to enhance production; the model was then validated. This model allowed the determination of the optimal values of all operational variables included: batch and perfusion duration and perfusion rate. The highest productivity (2682 virus cm−3 h−1) was obtained under the following conditions: batch at 37 °C for 53 h followed by perfusion at 32 °C for 23 h with a perfusion rate of 0.107 h−1. This value was 3.5‐fold higher than the best result obtained in batch cultures for the same conditions of titer and quality. A sensitivity analysis of the parameters showed that the parameters that affect most the final productivity depend on the bioreaction phase: cell growth in batch culture and production and product degradation in perfusion culture. In part 2 of this paper, this model is extended to the first step of downstream processing, and the addition of further steps to the process is discussed in order to achieve global process optimization.
Bibliography:	istex:225C91BBDB4A5DA696FFACFC57319C1B95824213 ArticleID:BTPR9901466 ark:/67375/WNG-27RWNFSZ-W ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	8756-7938 1520-6033
DOI:	10.1021/bp9901466