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  • Open Access

Production of biologically active recombinant human factor H in Physcomitrella

Authors

  • Annette Büttner-Mainik,

    1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany
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  • Juliana Parsons,

    1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany
    2. Centre for Biological Signalling Studies (BIOSS), Freiburg, Germany
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  • Hanna Jérôme,

    1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany
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  • Andrea Hartmann,

    1. Department Infection Biology, Leibniz Institute for Natural Product Research and Infectionbiology – Hans Knoell Institute, Jena, Germany
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  • Stephanie Lamer,

    1. Core Facility Proteomics, Centre for Systems Biology (ZBSA), Freiburg, Germany
    2. Feiburg Initiative for Systems Biology (FRISYS), Freiburg, Germany
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  • Andreas Schaaf,

    1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany
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    • Current address: greenovation Biotech, 79111 Freiburg, Germany.

  • Andreas Schlosser,

    1. Core Facility Proteomics, Centre for Systems Biology (ZBSA), Freiburg, Germany
    2. Feiburg Initiative for Systems Biology (FRISYS), Freiburg, Germany
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  • Peter F. Zipfel,

    1. Department Infection Biology, Leibniz Institute for Natural Product Research and Infectionbiology – Hans Knoell Institute, Jena, Germany
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  • Ralf Reski,

    1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany
    2. Centre for Biological Signalling Studies (BIOSS), Freiburg, Germany
    3. Feiburg Initiative for Systems Biology (FRISYS), Freiburg, Germany
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  • Eva L. Decker

    Corresponding author
    1. Plant Biotechnology, Faculty of Biology, University of Freiburg, Freiburg, Germany
    2. Centre for Biological Signalling Studies (BIOSS), Freiburg, Germany
      (email eva.decker@biologie.uni-freiburg.de)
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(email eva.decker@biologie.uni-freiburg.de)

Summary

The human complement regulatory serum protein factor H (FH) is a promising future biopharmaceutical. Defects in the gene encoding FH are associated with human diseases like severe kidney and retinal disorders in the form of atypical haemolytic uremic syndrome (aHUS), membranoproliferative glomerulonephritis II (MPGN II) or age-related macular degeneration (AMD). There is a current need to apply intact full-length FH for the therapy of patients with congenital or acquired defects of this protein. Application of purified or recombinant FH (rFH) to these patients is an important and promising approach for the treatment of these diseases. However, neither protein purified from plasma of healthy individuals nor recombinant protein is currently available on the market. Here, we report the first stable expression of the full-length human FH cDNA and the subsequent production of this glycoprotein in a plant system. The moss Physcomitrella patens perfectly suits the requirements for the production of complex biopharmaceuticals as this eukaryotic system not only offers an outstanding genetical accessibility, but moreover, proteins can be produced safely in scalable photobioreactors without the need for animal-derived medium compounds. Transgenic moss lines were created, which express the human FH cDNA and target the recombinant protein to the culture supernatant via a moss-derived secretion signal. Correct processing of the signal peptide and integrity of the moss-produced rFH were verified via peptide mapping by mass spectrometry. Ultimately, we show that the rFH displays complement regulatory activity comparable to FH purified from plasma.

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