title: Purification of Microbially Expressed Recombinant Proteins via a Dual ELP Split Intein System.
abstract: Fusions of elastin-like peptide (ELP) purification tags and self-cleaving inteins provide a powerful platform for purifying tagless recombinant proteins without the need for conventional packed-bed columns. A drawback to this method has been premature cleaving of the ELP tag during expression, before the purification procedure can take place. Here we demonstrate a split-intein method, where the self-cleaving intein is divided into two inactive segments during expression and purification. Spontaneous assembly of the purified intein segments then restores self-cleaving activity to deliver the tagless target protein.
authors.affiliation: Biodesign Institute at Arizona State University, Tempe, AZ, USA.
authors.forename: Changhua
authors.initials: C
authors.lastname: Shi
authors.affiliation: Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Tzu-Chiang
authors.initials: TC
authors.lastname: Han
authors.affiliation: Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH, 43210, USA. wood.750@osu.edu.
authors.forename: David W
authors.initials: DW
authors.lastname: Wood
mesh_terms: D004549:Elastin / Q000737:chemistry / Q000235:genetics; D047668:Inteins; D010455:Peptides / Q000737:chemistry / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; D004549:Elastin
keywords: Elastin like protein; Intein; Non-chromatographic purification; Recombinant protein purification; Split intein; Trans cleavage