Benchabane Meriem, Rivard Daniel, Girard Cécile, Michaud Dominique
Meriem Département de phytologie, Université Laval, Québec, Canada.
Methods Mol Biol. 2009;483:265-73. doi: 10.1007/978-1-59745-407-0_15.
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.
我们描述了一种将重组蛋白酶抑制剂用作从转基因植物组织中提取的临床有用蛋白质稳定剂的通用方法。首先描述了一种评估转基因植物粗蛋白提取物中异源蛋白整体(不)稳定性的程序。然后给出了逐步方案,用于选择和使用在提取过程中抑制宿主植物蛋白酶的伴侣蛋白酶抑制剂。这种策略再现了通常添加到蛋白质提取介质中的低分子量蛋白酶抑制剂的蛋白质稳定作用,不需要外源添加此类昂贵且通常有毒的化合物。它还具有本质上可根据处理的生物量进行扩展的优点。