Summary
We describe a general approach for the use of recombinant protease inhibitors as stabilizing agents for clinically useful proteins extracted from transgenic plant tissues. A procedure is first described to assess the overall (in)stability of heterologous proteins in transgenic plant crude protein extracts. Step-by-step protocols are then presented for the choice and use of companion protease inhibitors inhibiting the host plant proteases during extraction. This strategy, that reproduces the protein-stabilizing effect of low-molecular-weight protease inhibitors often added to protein extraction media, does not require the exogenous addition of such expensive and often toxic compounds. It also presents the advantage of being intrinsically scalable to the amount of biomass processed.
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Acknowledgments
This work was supported by a Discovery grant from the Natural Science and Engineering Research Council (NSERC) of Canada to D. Michaud. M. Benchabane was the recipient of an NSERC doctoral scholarship.
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Benchabane, M., Rivard, D., Girard, C., Michaud, D. (2009). Companion Protease Inhibitors to Protect Recombinant Proteins in Transgenic Plant Extracts. In: Faye, L., Gomord, V. (eds) Recombinant Proteins From Plants. Methods in Molecular Biology™, vol 483. Humana Press. https://doi.org/10.1007/978-1-59745-407-0_15
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DOI: https://doi.org/10.1007/978-1-59745-407-0_15
Publisher Name: Humana Press
Print ISBN: 978-1-58829-978-9
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