Retroviral vector production using suspension-adapted 293GPG cells in a 3L acoustic filter-based perfusion bioreactor

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1002/bit.20947
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Name affiliation
  1. National Research Council of Canada. NRC Biotechnology Research Institute
FormatText, Article
Journal titleBiotechnology and Bioengineering
ISSN0006-3592
1097-0290
Volume95
Issue4
Pages653–660
Subjectretroviral vectors; HEK‐293 cells; suspension culture; bioreactors; perfusion culture; culture condition
Abstract
Recombinant retroviruses are now an established tool for gene delivery. Presently they are mainly produced using adherent cells. However, due to the restrictive nature of adherent cell culture, this mode of production is hampered by low cell‐specific productivity and small production units. The large‐scale production of retroviral vectors could benefit from the adaptation of retrovirus packaging cell lines to suspension culture. Here, we describe the ability of a 293 packaging cell line to produce retroviral vectors in suspension culture at high titer. Adherent 293GPG cells, producing a Moloney Murine Leukemia Virus (MoMLV) retrovirus vector pseudotyped with the vesicular stomatitis virus G (VSVG) envelope protein and expressing a TK‐GFP fusion protein, were adapted to suspension culture in calcium‐free DMEM. At a cell density similar to adherent cell culture, the suspension culture produced retroviral vector consistently in the range of 1 × 107 infectious viral particles/mL (IVP/mL), with a specific productivity threefold higher than adherent culture. Furthermore, at the same medium replacement frequency, the suspension producer cells could be cultured at higher density than their adherent counterparts, which resulted in virus titer of 3–4 × 107 IVP/mL at 11.0 × 106 cells/mL. This corresponds to a 10‐fold increase in viral concentration compared to adherent cells. The capacity to up scale the retroviral vector production was also demonstrated by performing a 2 VVD perfusion culture for 9 days in a 3L Chemap bioreactor. The combination of suspension and perfusion led to a 20‐fold increase in maximum virus productivity compared to the adherent culture.
Publication date
PublisherWiley
NoteDA - 20061003IS - 0006-3592 (Print)LA - engPT - Journal ArticlePT - Research Support, Non-U.S. Gov'tRN - 0 (Viral Envelope Proteins)SB - IM
LanguageEnglish
Peer reviewedYes
NRC numberNRC-BRI-47776
NPARC number3539682
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Record identifier6eb1e460-25e5-4214-9bfe-6c7eb67971eb
Record created2009-03-01
Record modified2020-04-22
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