Proteolytic Affinity Tag Cleavage

Abstract

Here, we present protocols describing the use of the dipeptidyl-aminopeptidase-1 (DPP1, DAPase) exoprotease-based TAGZyme system and the endoprotease, Factor Xa. Both enable the recovery of proteins free of any amino acids encoded by the vector and/or protease recognition site. They also provide the possibility of removing the proteases from the preparation of the target protein by a simple subtractive chromatography step. TAGZyme enzymes contain an uncleavable His tag for removal by Immobilized Metal Ion Affinity Chromatography (IMAC). Factor Xa can be removed using Xa Removal Resin.

Keywords: DAPase enzyme; Exoproteolytic cleavage; GST fusion protein; Glutathione S-Transferase tag (GST tag); Immobilized metal ion affinity chromatography (IMAC); Qcyclase enzymes; TAGZyme; Xa removal resin.