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Baculovirus expression system scaleup by perfusion of high-density Sf-9 cell cultures

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1002/bit.260430907
AuteurRechercher : 1; Rechercher : 1; Rechercher : 1; Rechercher : 1
Affiliation
  1. Conseil national de recherches du Canada. Institut de recherche en biotechnologie du CNRC
FormatTexte, Article
Sujetbaculovirus; recombinant proteins; Sf-9 insect cells; perfusion; tangential filtration
Résumé
A perfusion system based on a 4-L stirred tank bioreactor and a custom-designed tangential (cross-flow) filter was assembled to realize a scaleup of the Baculovirus Expression Vector System (BEVS). When perfused with 1 to 1.5 vol/day, Spodoptera frugiperda (Sf-9) insect cell cultures grew from 4 x l0⁶ to 15 x l0⁶ cells/mL over 3 to 4 days. The possibility of maintaining high specific production of recombinant VP6 protein (from bovine rotavirus) after baculovirus infection of the high-density cultures was then assessed. The process consisted of a growth phase in TNMFH + 10% FBS, followed by infection with Bac-BRV6L recombinant baculovirus and a shift to a low-serum (0 to 1%) medium for perfusion during the production phase. Multiple runs were executed, each including a battery of shaker flask controls at various cell densities and serum concentrations. On average, specific rVP6 production in the bioreactor amounted to 76% of that found in 20-mL shaker cultures simulating the bioreactor's high cell density, low serum concentration, and medium renewal rate. Mechanical stress generated by cell/medium separation in the perfusion process reduced cell growth rate but had minimal effect on rVP6 production. Our results also indicated that serum concentration during the infection phase affected the rVP6 specific production in a cell density-dependent fashion. Although the feasibility of the cell density scale up was demonstrated, optimization is still needed to achieve a truly cost-effective process.
Date de publication
Dans
  • Biotechnology and Bioengineering 43, nº 9 (15 avril 1994) : 881–891.
Langueanglais
Numéro du CNRC38479
Numéro NPARC12338926
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Identificateur de l’enregistrementa37c7c0a-bad8-440d-972a-b7f44abf9ba4
Enregistrement créé2009-09-11
Enregistrement modifié2020-04-27
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