Oriented immobilization of the tobacco etch virus protease for the cleavage of fusion proteins

► We immobilize Streptag II-TEV protease in oriented manner on streptavidin matrix. ► Affinity immobilized enzyme retains more than 80% of its activity. ► Streptag II/streptavidin based immobilization improves 2-4 times enzyme stability. ► Affinity immobilization offers reuse of TEV protease during long periods. ► TEV proteolytic column shortens cleavage process of streptagged fusion proteins.