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Influence of growth temperature on the production of antibody Fab fragments in different microbes: A host comparative analysis

Authors

  • Martin Dragosits,

    1. Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
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  • Gianni Frascotti,

    1. Dept. of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
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    • Martin Dragosits, Gianni Frascotti, Lise Bernard-Granger, Felícitas Vázquez, Maria Giuliani, and Kristin Baumann contributed equally to this study

  • Lise Bernard-Granger,

    1. VTT Technical Research Centre, Espoo, Finland
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    • Martin Dragosits, Gianni Frascotti, Lise Bernard-Granger, Felícitas Vázquez, Maria Giuliani, and Kristin Baumann contributed equally to this study

  • Felícitas Vázquez,

    1. Institut de Biotecnologia i de Biomedicina and Dept. of Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Barcelona, Spain
    2. CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, Barcelona 08193, Spain
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    • Martin Dragosits, Gianni Frascotti, Lise Bernard-Granger, Felícitas Vázquez, Maria Giuliani, and Kristin Baumann contributed equally to this study

  • Maria Giuliani,

    1. Dept. of Organic Chemistry and Biochemistry, Università di Napoli Federico II, Naples, Italy
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    • Martin Dragosits, Gianni Frascotti, Lise Bernard-Granger, Felícitas Vázquez, Maria Giuliani, and Kristin Baumann contributed equally to this study

  • Kristin Baumann,

    1. Dept. of Chemical Engineering, Escola Tècnica Superior d'Enginyeria, Universitat Autònoma de Barcelona, Barcelona, Spain
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    • Martin Dragosits, Gianni Frascotti, Lise Bernard-Granger, Felícitas Vázquez, Maria Giuliani, and Kristin Baumann contributed equally to this study

  • Escarlata Rodríguez-Carmona,

    1. Institut de Biotecnologia i de Biomedicina and Dept. of Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Barcelona, Spain
    2. CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, Barcelona 08193, Spain
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  • Jaana Tokkanen,

    1. VTT Technical Research Centre, Espoo, Finland
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  • Ermenegilda Parrilli,

    1. Dept. of Organic Chemistry and Biochemistry, Università di Napoli Federico II, Naples, Italy
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  • Marilyn G. Wiebe,

    1. VTT Technical Research Centre, Espoo, Finland
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  • Renate Kunert,

    1. Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
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  • Michael Maurer,

    1. Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
    2. School of Bioengineering, University of Applied Sciences FH-Campus, Wien, Austria
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  • Brigitte Gasser,

    1. Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
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  • Michael Sauer,

    1. Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
    2. School of Bioengineering, University of Applied Sciences FH-Campus, Wien, Austria
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  • Paola Branduardi,

    1. Dept. of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
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  • Tiina Pakula,

    1. VTT Technical Research Centre, Espoo, Finland
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  • Markku Saloheimo,

    1. VTT Technical Research Centre, Espoo, Finland
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  • Merja Penttilä,

    1. VTT Technical Research Centre, Espoo, Finland
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  • Pau Ferrer,

    1. Dept. of Chemical Engineering, Escola Tècnica Superior d'Enginyeria, Universitat Autònoma de Barcelona, Barcelona, Spain
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  • Maria Luisa Tutino,

    1. Dept. of Organic Chemistry and Biochemistry, Università di Napoli Federico II, Naples, Italy
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  • Antonio Villaverde,

    1. Institut de Biotecnologia i de Biomedicina and Dept. of Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Barcelona, Spain
    2. CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Bellaterra, Barcelona 08193, Spain
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  • Danilo Porro,

    1. Dept. of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy
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  • Diethard Mattanovich

    Corresponding author
    1. Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
    2. School of Bioengineering, University of Applied Sciences FH-Campus, Wien, Austria
    • Dept. of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria
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Errata

This article is corrected by:

  1. Errata: Erratum Volume 28, Issue 4, 1114, Article first published online: 23 July 2012

Abstract

Microorganisms encounter diverse stress conditions in their native habitats but also during fermentation processes, which have an impact on industrial process performance. These environmental stresses and the physiological reactions they trigger, including changes in the protein folding/secretion machinery, are highly interrelated. Thus, the investigation of environmental factors, which influence protein expression and secretion is still of great importance. Among all the possible stresses, temperature appears particularly important for bioreactor cultivation of recombinant hosts, as reductions of growth temperature have been reported to increase recombinant protein production in various host organisms. Therefore, the impact of temperature on the secretion of proteins with therapeutic interest, exemplified by a model antibody Fab fragment, was analyzed in five different microbial protein production hosts growing under steady-state conditions in carbon-limited chemostat cultivations. Secretory expression of the heterodimeric antibody Fab fragment was successful in all five microbial host systems, namely Saccharomyces cerevisiae, Pichia pastoris, Trichoderma reesei, Escherichia coli and Pseudoalteromonas haloplanktis. In this comparative analysis we show that a reduction of cultivation temperature during growth at constant growth rate had a positive effect on Fab 3H6 production in three of four analyzed microorganisms, indicating common physiological responses, which favor recombinant protein production in prokaryotic as well as eukaryotic microbes. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2011

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