Hedhammar My, Alm Tove, Gräslund Torbjörn, Hober Sophia
Department of Biotechnology, Royal Institute of Technology, AlbaNova University Center, 106 91 Stockholm, Sweden.
Biotechnol J. 2006 Feb;1(2):187-96. doi: 10.1002/biot.200500023.
A strategy for purification of inclusion body-forming proteins is described, in which the positively charged domain Z(basic) is used as a fusion partner for capture of denatured proteins on a cation exchange column. It is shown that the purification tag is selective under denaturing conditions. Furthermore, the new strategy for purification of proteins from inclusion bodies is compared with the commonly used method for purification of His(6)-tagged inclusion body proteins. Finally, the simple and effective means of target protein capture provided by the Z(basic) tag is further successfully explored for solid-phase refolding. This procedure has the inherited advantage of combining purification and refolding in one step and offers the advantage of eluting the concentrated product in a suitable buffer.
本文描述了一种纯化包涵体形成蛋白的策略，其中带正电荷的结构域Z（碱性）用作融合伴侣，用于在阳离子交换柱上捕获变性蛋白。结果表明，该纯化标签在变性条件下具有选择性。此外，将这种从包涵体中纯化蛋白质的新策略与常用的纯化His(6)标签包涵体蛋白的方法进行了比较。最后，进一步成功探索了Z（碱性）标签提供的简单有效的靶蛋白捕获方法用于固相重折叠。该方法具有一步结合纯化和重折叠的固有优势，并具有在合适缓冲液中洗脱浓缩产物的优点。
