title: 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.
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Helena
authors.initials: H
authors.lastname: Block
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Barbara
authors.initials: B
authors.lastname: Maertens
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Anne
authors.initials: A
authors.lastname: Spriestersbach
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Kubicek
authors.affiliation: QIAGEN GmbH, Research and Development, Qiagenstrasse 1, 40724 Hilden, Germany. Electronic address: frank.schaefer@qiagen.com.
authors.forename: Frank
authors.initials: F
authors.lastname: Schäfer
mesh_terms: D000818:Animals; D001688:Biological Products / Q000737:chemistry; D002021:Buffers; D020789:Cathepsin C / Q000737:chemistry; D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D020689:Exopeptidases / Q000737:chemistry*; D015951:Factor Xa / Q000737:chemistry; D005982:Glutathione Transferase / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions; D008670:Metals / Q000737:chemistry; D059748:Proteolysis; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D001688:Biological Products; D002021:Buffers; D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D006639:Histidine; D005982:Glutathione Transferase; D020689:Exopeptidases; D020789:Cathepsin C; D015951:Factor Xa
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