Abstract
Retroviral vectors are involved in more than 25% of the gene therapy trials, being the pioneering vector in this field. The production of retroviral vectors still poses some challenges, mainly because of the relatively low cell specific productivity and the low vector stability. Having clinical applications in mind, it is clear that robust production and purification processes are necessary, along with a proper vector characterization. The determination of vector titer in terms of infectious particles and transduction efficiency, combined with product quality analysis in terms of total particles and proteins present in the final preparation, are essential for a complete assessment and definition of the final product specifications.
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References
Andreadis, S. T., Roth, C. M., Le Doux, J. M., Morgan, J. R., and Yarmush, M. L. (1999) Large-scale processing of recombinant retroviruses for gene therapy. Biotechnol. Prog. 15, 1–11.
Mountain, A. (2000) Gene therapy: the first decade. Trends Biotechnol. 18, 119–128.
Thomas, C. E., Ehrhardt, A., and Kay, M. A. (2003) Progress and problems with the use of viral vectors for gene therapy. Nat. Rev. Genet. 4, 346–358.
Relph, K., Harrington, K., and Pandha, H. (2004) Recent developments and current status of gene therapy using viral vectors in the United Kingdom. BMJ 329, 839–842.
Chuck, A. S. and Palsson, B. O. (1996) Consistent and high rates of gene transfer can be obtained using flow-through transduction over a wide range of retroviral titers. Hum. Gene Ther. 7, 743–750.
McTaggart, S. and Al-Rubeai, M. (2000) Effects of culture parameters on the production of retroviral vectors by a human packaging cell line. Biotechnol. Prog. 16, 859–865.
Carmo, M., Peixoto, C., Coroadinha, A. S., Alves, P. M., Cruz, P. E., and Carrondo, M. J. (2004) Quantitation of MLV-based retroviral vectors using real-time RT-PCR. J. Virol. Methods 119, 115–119.
Le Doux, J. M., Davis, H. E., Morgan, J. R., and Yarmush, M. L. (1999) Kinetics of retrovirus production and decay. Biotechnol. Bioeng. 63, 654–662.
Pizzato, M., Merten, O. W., Blair, E. D., and Takeuchi, Y. (2001) Development of a suspension packaging cell line for production of high titre, serum-resistant murine leukemia virus vectors. Gene Ther. 8, 737–745.
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© 2007 Humana Press Inc., Totowa, NJ
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Cruz, P.E., Carmo, M., Rodrigues, T., Alves, P. (2007). Retrovirus Production and Characterization. In: Pörtner, R. (eds) Animal Cell Biotechnology. Methods in Biotechnology, vol 24. Humana Press. https://doi.org/10.1007/978-1-59745-399-8_22
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DOI: https://doi.org/10.1007/978-1-59745-399-8_22
Publisher Name: Humana Press
Print ISBN: 978-1-58829-660-3
Online ISBN: 978-1-59745-399-8
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