Large-scale production of lentiviral vectors using multilayer cell factories
Lentiviral-mediated gene therapy has been proposed for the treatment of a range of diseases, and due to its genome integration properties, it offers the potential for long-lasting benefit from a once-off treatment. Production methods for pre-clinical studies in animal models, and ultimately for human clinical trials, must be capable of producing large quantities of high-quality lentiviral vector in an efficient and cost-effective manner. We report here a medium-scale method (from 1.5 L to 6 L of vector supernatant) for lentiviral vector production in adherent cell cultures using the NUNC™ EasyFill™ Cell Factory™ from Thermo Fisher Scientific. Downstream purification uses a Mustang Q XT5 anion exchange capsule from Pall, and an ultracentrifugation step to concentrate the vector. This method is capable of producing lentiviral vector with concentrated titres of 108–109 TU/ml, with reduced manual handling compared to single monolayer flask methods.
calcium phosphate, cell factories, lentiviral vector, Mustang Q, ultracentrifugation
Citation: Rout-Pitt N, McCarron A, McIntyre C, Parsons D, Donnelley M. Large-scale production of lentiviral vectors using multilayer cell factories. J Biol Methods 2018;5(2):e90. doi: 10.14440/jbm.2018.236
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