Companion protease inhibitors to protect recombinant proteins in transgenic plant extracts

Abstract

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. © 2009 Humana Press.