Z_basic: A Purification Tag for Selective Ion-Exchange Recovery

A positively charged protein domain, denoted Z_basic, can be used as a general purification tag for purification of recombinantly produced target proteins by cation-exchange chromatography. The Z_basic domain is constructed from the Protein A-derived Z-domain, and engineered to be highly charged, which allows selective capture on a cation exchanger at physiological pH values. Moreover, Z_basic is selective also under denaturing conditions and can be used for purification of proteins solubilized from inclusion bodies. Z_basic can then be used as a flexible linker to the cation-exchanger resin, and thereby allows solid-phase refolding of the target protein.Herein, protocols for purification of soluble Z_basic-tagged fusion proteins , as well as for integrated purification and solid-phase refolding of insoluble fusion proteins , are described. In addition, a procedure for enzymatic tag removal and recovery of native target protein is outlined.