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Improving yield of a recombinant biologic in a Brassica hairy root manufacturing process
  • +5
  • Noemi Gutierrez-Valdes,
  • Suvi Häkkinen,
  • Camille Lemasson,
  • Jonas de Groot,
  • Jean-Pierre Ele-Ekouna,
  • Marina Guillet,
  • Florian Cardon,
  • Anneli Ritala
Noemi Gutierrez-Valdes
Teknologian tutkimuskeskus VTT Oy

Corresponding Author:[email protected]

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Suvi Häkkinen
Teknologian tutkimuskeskus VTT Oy
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Camille Lemasson
Samabriva Amiens France
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Jonas de Groot
Teknologian tutkimuskeskus VTT Oy
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Jean-Pierre Ele-Ekouna
BIOPI Biologie Des Plantes Et Innovation Amiens France
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Marina Guillet
Samabriva Amiens France
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Florian Cardon
Samabriva Amiens France
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Anneli Ritala
Teknologian tutkimuskeskus VTT Oy
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Hairy root systems have proven to be a viable alternative for recombinant protein production. For recalcitrant proteins, maximizing the productivity of hairy root cultures is essential. The aim of this study was to optimize a Brassica rapa rapa hairy root process for secretion of α-L-iduronidase (IDUA), a biologic of medical value. The process was first optimized with hairy roots expressing eGFP. For the biomass optimization, the highest biomass yields were achieved in modified Gamborg B5 culture medium. For the secretion induction, the optimized secretion media was obtained with additives (1.5g/l PVP + 1mg/l 2,4-D + 20.5g/l KNO 3) resulting in 3.4 fold eGFP secretion when compared to the non-induced control. These optimized conditions were applied to the IDUA-expressing hairy root clone, confirming that the highest yields of secreted IDUA occurred when using the already defined additive combination. The functionality of the IDUA protein, secreted and intracellular, was confirmed with an enzymatic activity assay. A >150-fold increase of the IDUA activity was observed using an optimized secretion medium, compared with a non-induced medium. We have proven that our B. rapa rapa hairy root system can be harnessed to secrete recalcitrant proteins, illustrating the high potential of hairy roots in plant molecular farming.
28 Feb 2022Submitted to Biotechnology and Bioengineering
28 Feb 2022Submission Checks Completed
28 Feb 2022Assigned to Editor
11 Mar 2022Reviewer(s) Assigned
11 Apr 2022Editorial Decision: Revise Major
11 Apr 2022Review(s) Completed, Editorial Evaluation Pending
20 May 20221st Revision Received
24 May 2022Submission Checks Completed
24 May 2022Assigned to Editor
07 Jun 2022Review(s) Completed, Editorial Evaluation Pending
07 Jun 2022Editorial Decision: Accept
Oct 2022Published in Biotechnology and Bioengineering volume 119 issue 10 on pages 2831-2841. 10.1002/bit.28178