Summary
Gene therapy is becoming increasingly relevant for the treatment of prominent human diseases. Viral vectors are currently used in more than 50% of the gene therapy clinical trials, most of them aimed at cancer diseases. Clearly, the increasing needs of high-quality viral preparations required the elimination of process bottlenecks, streamlining the development of the viral into a real-world clinical tool . Virus production for clinical gene therapy can be a limiting step because many virus generation protocols rely on labor-intensive, bench-scale methods; robust, cost-effective strategies for the delivery of clinical-grade viruses are thus essential for the future of gene therapy. A comprehensive picture of key aspects on the integration of upstream and downstream processing is addressed in this chapter, by describing the case study of recombinant budded baculoviruses for gene therapy; scalable methods are described in detail as well as mandatory characterization techniques for a proper and complete quality assessment of the viral vectors.
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Notes
- 1.
“Viral quality” (total viruses/infective viruses ratio) or “viral titer” can be used to define the harvest optimum, depending on the process specificities and the strategy of optimization available.
- 2.
Cells previously transfected thereby becoming capable of permanently generating a designed viral vector.
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Acknowledgments
We thank Marcos Sousa for the expertise on the upstream processing and Dr. Uwe Gottschalk from Sartorius Stedim Biotech (Göttingen, Germany) for providing the Sartobind membrane adsorber units and the ultrafiltration device. Financial support from the European Commission (Baculogenes, LSH-2005–1.4.4.6) and the Portuguese Fundação para a Ciência e Tecnologia (SFRH/BD/31257/2006) is gratefully acknowledged.
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Vicente, T., Peixoto, C., Carrondo, M.J., Alves, P.M. (2009). Virus Production for Clinical Gene Therapy. In: Walther, W., Stein, U. (eds) Gene Therapy of Cancer. Methods in Molecular Biology™, vol 542. Humana Press. https://doi.org/10.1007/978-1-59745-561-9_24
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