title: Recombinant protein purification by self-cleaving elastin-like polypeptide fusion tag.
abstract: This unit presents a rapid and simple method for the nonchromatographic purification of recombinant proteins expressed in E. coli. This method relies on a thermally responsive elastin-like polypeptide (ELP) tag, where the tagged protein is precipitated using a mild temperature shift. The tag is then induced to self-cleave by a mild pH shift and is subsequently removed by a final thermal precipitation. The result is a purified native protein target, without the requirement for affinity apparatus or protease removal of the tag. This protocol describes the required cloning methods to insert a given target into the expression vector, as well as the general method for purifying the resulting expressed protein.
authors: Princeton University, Princeton, New Jersey.
authors: Wan-Yi
authors: WY
authors: Wu
authors: Princeton University, Princeton, New Jersey.
authors: Baley A
authors: BA
authors: Fong
authors: Princeton University, Princeton, New Jersey.
authors: Allison G
authors: AG
authors: Gilles
authors: Princeton University, Princeton, New Jersey.
authors: David W
authors: DW
authors: Wood
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D004549:Elastin / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011506:Proteins; D011993:Recombinant Fusion Proteins; D004549:Elastin