Virus-like particle production at low multiplicities of infection with the baculovirus insect cell system
The baculovirus insect cell expression system (BEVS) was used for the production of self‐forming Porcine parvovirus‐like particles (VLPs) in serum‐free medium. A low multiplicity of infection (MOI) strategy was used to overcome an extra virus amplification step, undesirable in industrial production,...
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Published in | Biotechnology and bioengineering Vol. 84; no. 2; pp. 245 - 253 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Hoboken
Wiley Subscription Services, Inc., A Wiley Company
20.10.2003
Wiley |
Subjects | |
Online Access | Get full text |
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Summary: | The baculovirus insect cell expression system (BEVS) was used for the production of self‐forming Porcine parvovirus‐like particles (VLPs) in serum‐free medium. A low multiplicity of infection (MOI) strategy was used to overcome an extra virus amplification step, undesirable in industrial production, and to minimize the virus passage effect. It was confirmed that the time of infection (TOI) and MOI are dependent variables. Higher cell densities were obtained at low MOIs, keeping a constant TOI; however, both volumetric and specific productivities were lower. In synchronous infection, at high MOI, the specific productivity decreased when the cells were infected in the late phase of growth. Product degradation due to cell lysis strongly influenced the optimal time of harvest (TOH). Time of harvest was found to be highly dependent on the MOI, and a direct relationship with the cell yield was obtained.
Analysis of the culture medium reveals that glutamine depletion occurs in the late phase of the growth. Supplementation of glutamine to uninfected cell cultures resulted in an increased cell yield. Its addition to cultures infected in the middle phase of the growth curve was also able to restore the productivity levels, but addition to cells in their stationary phase caused no observable effect on product expression. The study clearly shows that for a specific TOI it is not obvious what the correct MOI should be to obtain the best volumetric productivity. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 84: 245–253, 2003. |
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Bibliography: | Fundação para a Ciência e Tecnologia-Portugal European Commission - No. BIO4-CT98-0215; No. PRAXIS XXI/BD/16136/98 istex:5725C600B53BE9531ACD32941EB0A631320C15A3 ArticleID:BIT10773 ark:/67375/WNG-FST7MCT6-G ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 |
ISSN: | 0006-3592 1097-0290 |
DOI: | 10.1002/bit.10773 |