Open AccessElastin‐Like Polypeptides as a Purification Tag for Recombinant Proteins
Author(s) -
Hassouneh Wafa,
Christensen Trine,
Chilkoti Ashutosh
Publication year - 2010
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps0611s61
Subject(s) - recombinant dna , fusion protein , flag tag , elastin , chemistry , myc tag , protein purification , multiplex , fusion , chromatography , biochemistry , biology , bioinformatics , genetics , linguistics , philosophy , gene
This unit presents a recombinant protein purification method that employs an elastin‐like polypeptide (ELP) as a purification tag. ELPs undergo a sharp and reversible phase transition when heated above their lower critical solution temperature. ELPs retain this behavior when they are fused to a protein, and thereby provide a simple method to isolate a recombinant ELP fusion protein from cell contaminants by cycling the solution through the insoluble and soluble phase of the ELP fusion protein using a procedure that is termed Inverse Transition Cycling. This method does not require the use of chromatography, so it is cost‐effective, easy to scale up, and easy to multiplex. Curr. Protoc. Protein Sci . 61:6.11.1‐6.11.16. © 2010 by John Wiley & Sons, Inc.