293 Cell Cycle Synchronisation In Adenovirus Vector Production

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

As the market requirements for adenovirus vectors (AdV) increase, the maximisation of the virus titer per culture volume per unit time is a key requirement. However, despite the fact that 293 cells can grow up to 8 x 10(6) cell/mL in simple batch mode operations, for optimal AdV infection a maximum cell density of 1 x 10(6) cell/mL at infection time has usually been utilized due to the so called "cell density effect". In addition, AdV titer appears to be dependent upon cell cycle phase at the time of infection. To evaluate the dependence of AdV production upon cell cycle phase, 293 cells were chemically synchronised at each phase of the cell cycle; a 2.6-fold increase on AdV cell specific titer was obtained whet? the percentage of cells at the S phase of the cell cycle was increased from 36 to 47%; a mathematical equation was used to relate AdV cell specific productivities with cell synchronisation at the S phase using this data. To avoid the use of chemical inhibitors, a temperature shift strategy was also used for synchronisation at the S phase. S phase synchronisation was obtained by decreasing the culture temperature to 31 degrees C during 67 h and restoring it to 37 degrees C during 72 h. By using this strategy we were able to synchronise 57% of the population in the S phase of the cell cycle obtaining an increase of 7.3-fold on AdV cell specific titer after infection. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 235-243, 2009
Original languageUnknown
Pages (from-to)235-243
JournalBiotechnology Progress
Volume25
Issue number1
DOIs
Publication statusPublished - 1 Jan 2009

Keywords

    Cite this

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    title = "293 Cell Cycle Synchronisation In Adenovirus Vector Production",
    abstract = "As the market requirements for adenovirus vectors (AdV) increase, the maximisation of the virus titer per culture volume per unit time is a key requirement. However, despite the fact that 293 cells can grow up to 8 x 10(6) cell/mL in simple batch mode operations, for optimal AdV infection a maximum cell density of 1 x 10(6) cell/mL at infection time has usually been utilized due to the so called {"}cell density effect{"}. In addition, AdV titer appears to be dependent upon cell cycle phase at the time of infection. To evaluate the dependence of AdV production upon cell cycle phase, 293 cells were chemically synchronised at each phase of the cell cycle; a 2.6-fold increase on AdV cell specific titer was obtained whet? the percentage of cells at the S phase of the cell cycle was increased from 36 to 47{\%}; a mathematical equation was used to relate AdV cell specific productivities with cell synchronisation at the S phase using this data. To avoid the use of chemical inhibitors, a temperature shift strategy was also used for synchronisation at the S phase. S phase synchronisation was obtained by decreasing the culture temperature to 31 degrees C during 67 h and restoring it to 37 degrees C during 72 h. By using this strategy we were able to synchronise 57{\%} of the population in the S phase of the cell cycle obtaining an increase of 7.3-fold on AdV cell specific titer after infection. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 235-243, 2009",
    keywords = "replication, expression, density, arrest, recombinant, cell, 293, cycle, synchronisation, s-phase, fibroblasts, cells, system, propagation, culture, adenovirus, effect, protein, mammalian-cells, temperature, growth",
    author = "Carrondo, {Manuel Jos{\'e} Teixeira} and Alves, {Paula Maria}",
    year = "2009",
    month = "1",
    day = "1",
    doi = "10.1002/btpr.64",
    language = "Unknown",
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    pages = "235--243",
    journal = "Biotechnology Progress",
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    publisher = "ACS American Chemical Society",
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    }

    293 Cell Cycle Synchronisation In Adenovirus Vector Production. / Carrondo, Manuel José Teixeira; Alves, Paula Maria.

    In: Biotechnology Progress, Vol. 25, No. 1, 01.01.2009, p. 235-243.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - 293 Cell Cycle Synchronisation In Adenovirus Vector Production

    AU - Carrondo, Manuel José Teixeira

    AU - Alves, Paula Maria

    PY - 2009/1/1

    Y1 - 2009/1/1

    N2 - As the market requirements for adenovirus vectors (AdV) increase, the maximisation of the virus titer per culture volume per unit time is a key requirement. However, despite the fact that 293 cells can grow up to 8 x 10(6) cell/mL in simple batch mode operations, for optimal AdV infection a maximum cell density of 1 x 10(6) cell/mL at infection time has usually been utilized due to the so called "cell density effect". In addition, AdV titer appears to be dependent upon cell cycle phase at the time of infection. To evaluate the dependence of AdV production upon cell cycle phase, 293 cells were chemically synchronised at each phase of the cell cycle; a 2.6-fold increase on AdV cell specific titer was obtained whet? the percentage of cells at the S phase of the cell cycle was increased from 36 to 47%; a mathematical equation was used to relate AdV cell specific productivities with cell synchronisation at the S phase using this data. To avoid the use of chemical inhibitors, a temperature shift strategy was also used for synchronisation at the S phase. S phase synchronisation was obtained by decreasing the culture temperature to 31 degrees C during 67 h and restoring it to 37 degrees C during 72 h. By using this strategy we were able to synchronise 57% of the population in the S phase of the cell cycle obtaining an increase of 7.3-fold on AdV cell specific titer after infection. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 235-243, 2009

    AB - As the market requirements for adenovirus vectors (AdV) increase, the maximisation of the virus titer per culture volume per unit time is a key requirement. However, despite the fact that 293 cells can grow up to 8 x 10(6) cell/mL in simple batch mode operations, for optimal AdV infection a maximum cell density of 1 x 10(6) cell/mL at infection time has usually been utilized due to the so called "cell density effect". In addition, AdV titer appears to be dependent upon cell cycle phase at the time of infection. To evaluate the dependence of AdV production upon cell cycle phase, 293 cells were chemically synchronised at each phase of the cell cycle; a 2.6-fold increase on AdV cell specific titer was obtained whet? the percentage of cells at the S phase of the cell cycle was increased from 36 to 47%; a mathematical equation was used to relate AdV cell specific productivities with cell synchronisation at the S phase using this data. To avoid the use of chemical inhibitors, a temperature shift strategy was also used for synchronisation at the S phase. S phase synchronisation was obtained by decreasing the culture temperature to 31 degrees C during 67 h and restoring it to 37 degrees C during 72 h. By using this strategy we were able to synchronise 57% of the population in the S phase of the cell cycle obtaining an increase of 7.3-fold on AdV cell specific titer after infection. (C) 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 235-243, 2009

    KW - replication

    KW - expression

    KW - density

    KW - arrest

    KW - recombinant

    KW - cell

    KW - 293

    KW - cycle

    KW - synchronisation

    KW - s-phase

    KW - fibroblasts

    KW - cells

    KW - system

    KW - propagation

    KW - culture

    KW - adenovirus

    KW - effect

    KW - protein

    KW - mammalian-cells

    KW - temperature

    KW - growth

    U2 - 10.1002/btpr.64

    DO - 10.1002/btpr.64

    M3 - Article

    VL - 25

    SP - 235

    EP - 243

    JO - Biotechnology Progress

    JF - Biotechnology Progress

    SN - 8756-7938

    IS - 1

    ER -