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

  1. Get@NRC: Baculovirus expression system scaleup by perfusion of high-density Sf-9 cell cultures (Opens in a new window)
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Journal titleBiotechnology and Bioengineering
Pages881891; # of pages: 11
Subjectbaculovirus; recombinant proteins; Sf-9 insect cells; perfusion; tangential filtration
AbstractA 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.
Publication date
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedNo
NRC number38479
NPARC number12338926
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Record identifiera37c7c0a-bad8-440d-972a-b7f44abf9ba4
Record created2009-09-11
Record modified2016-05-09
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