Companion Protease Inhibitors for the In Situ Protection of Recombinant Proteins in Plants

  • Stéphanie Robert
  • Philippe V. Jutras
  • Moustafa Khalf
  • Marc-André D’Aoust
  • Marie-Claire Goulet
  • Frank Sainsbury
  • Dominique Michaud
Part of the Methods in Molecular Biology book series (MIMB, volume 1385)


We previously described a procedure for the use of plant protease inhibitors as “companion” accessory proteins to prevent unwanted proteolysis of clinically useful recombinant proteins in leaf crude protein extracts (Benchabane et al. Methods Mol Biol 483:265–273, 2009). Here we describe the use of these inhibitors for the protection of recombinant proteins in planta, before their extraction from leaf tissues. A procedure is first described involving inhibitors co-expressed along—and co-migrating—with the protein of interest in host plant cells. An alternative, single transgene scheme is then described involving translational fusions of the recombinant protein and companion inhibitor. These approaches may allow for a significant improvement of protein steady-state levels in leaves, comparable to yield improvements observed with protease-deficient strains of less complex protein expression hosts such as E. coli or yeasts.


Clinically useful recombinant proteins Heterologous protein expression Recombinant protein degradation Companion protease inhibitors Protein stabilization 


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Stéphanie Robert
    • 1
  • Philippe V. Jutras
    • 1
  • Moustafa Khalf
    • 1
  • Marc-André D’Aoust
    • 2
  • Marie-Claire Goulet
    • 1
  • Frank Sainsbury
    • 1
    • 3
  • Dominique Michaud
    • 1
  1. 1.Centre de Recherche et d’Innovation sur les VégétauxUniversité LavalQuébecCanada
  2. 2.Medicago Inc.QuébecCanada
  3. 3.Australian Institute for Bioengineering and NanotechnologyThe University of QueenslandSt LuciaAustralia

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