title: Self-cleavable stimulus responsive tags for protein purification without chromatography.
abstract: A simple method to purify recombinant proteins is described by fusing a target protein with an intein and an elastin-like polypeptide that only requires NaCl, dithiothreitol, and a syringe filter to isolate the target protein from Escherichia coli lysate. This tripartite fusion system enables rapid isolation of the target protein without the need for affinity chromatography for purification or proteases for cleavage of the target protein from the fusion. The elastin-like polypeptide tag imparts reversible phase transition behavior to the tripartite fusion so that the fusion protein can be selectively aggregated in cell lysate by the addition of NaCl. The aggregates are isolated by microfiltration and resolubilized by reversal of the phase transition in low ionic strength buffer. After resolubilizing the fusion protein, the intein is activated to cleave the target protein from the elastin-intein tag, and the target protein is then isolated from the elastin-intein fusion by an additional phase transition cycle.
authors.affiliation: Department of Chemical Engineering, McMaster University, 1280 Main Street West JHE-377, Hamilton, Ontario L8S 4L7, Canada.
authors.forename: Xin
authors.initials: X
authors.lastname: Ge
authors.forename: Daniel S C
authors.initials: DS
authors.lastname: Yang
authors.forename: Kimberly
authors.initials: K
authors.lastname: Trabbic-Carlson
authors.forename: Bumjoon
authors.initials: B
authors.lastname: Kim
authors.forename: Ashutosh
authors.initials: A
authors.lastname: Chilkoti
authors.forename: Carlos D M
authors.initials: CD
authors.lastname: Filipe
mesh_terms: D002845:Chromatography; D004549:Elastin / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D005374:Filtration; D008832:Microchemistry; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins; D004549:Elastin