Large-scale production of lentiviral vectors using multilayer cell factories

Authors

  • Nathan Rout-Pitt Women's and Children's health Network, Respiratory and Sleep Medicine; University of Adelaide, Robinson Research Institute; University of Adelaide, Adelaide Medical School
  • Alexandra McCarron Women's and Children's health Network, Respiratory and Sleep Medicine; University of Adelaide, Robinson Research Institute; University of Adelaide, Adelaide Medical School
  • Chantelle McIntyre Women's and Children's health Network, Respiratory and Sleep Medicine; University of Adelaide, Robinson Research Institute; University of Adelaide, Adelaide Medical School
  • David Parsons Women's and Children's health Network, Respiratory and Sleep Medicine; University of Adelaide, Robinson Research Institute; University of Adelaide, Adelaide Medical School
  • Martin Donnelley Women's and Children's health Network, Respiratory and Sleep Medicine; University of Adelaide, Robinson Research Institute; University of Adelaide, Adelaide Medical School

DOI:

https://doi.org/10.14440/jbm.2018.236

Keywords:

calcium phosphate, cell factories, lentiviral vector, Mustang Q, ultracentrifugation

Abstract

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 NUNCâ„¢ EasyFillâ„¢ Cell Factoryâ„¢ 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.

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Published

2018-04-10

How to Cite

1.
Rout-Pitt N, McCarron A, McIntyre C, Parsons D, Donnelley M. Large-scale production of lentiviral vectors using multilayer cell factories. J Biol Methods [Internet]. 2018Apr.10 [cited 2022Aug.11];5(2):e90. Available from: https://jbmethods.org/jbm/article/view/236

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Section

Protocols