5.1.1 In such cases where AAV vectors are not ideal choices for gene therapy or ex vivo gene-modified cell therapies, lentiviral vectors are used. They typically encounter the same manufacturing hurdles as AAV, but the majority of LV batch requirements are for autologous cell therapies, so the total viral quantities needed may be smaller. The current generation of LV manufacturing requires four plasmids, as described in Figure 12.
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5.2.1 Transfection of multiple plasmids into a single cell to form LV vectors is similar, if not more complex, than transfection for AAV vectors. Likewise, there is great need for capable transfection agents that support more efficient, robust, and reproducible large-scale transfection and afford increased viral vector yields while maintaining compliance from quality, safety, and regulatory perspectives.
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5.3.1 The challenges to scalable upstream manufacturing of LV vectors are similar to those discussed for AAV vectors. Processes tend to be product-specific according to the gene of interest, typically take 3–5 weeks, and include expanding the seed train, inoculating the terminal reactor, and cell culture.[94]
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5.4.1 Because LV vectors for use in the production of CAR-T cell therapies were not intended for direct administration to humans but for the modification of cells that would be expanded and then administered to the patient, the first CAR-T products did not utilize a traditional DSP. Most products were purified under sterile filtration (0.2-micron filter).
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