title: A non-chromatographic protein purification strategy using Src 3 homology domains as generalized capture domains.
abstract: Protein purification using inverse phase transition of elastin-like polypeptide (ELP) domains is a useful alternative to chromatography. Genetic fusions of ELP domains to various proteins have the ability to reversibly transition between soluble monomers and micron-sized aggregates and this has been used to selectively purify many ELP fusions. Affinity domains can enhance this technology by using specific protein binding domains to enable ELP mediated affinity capture (EMAC) of proteins of interest (POI) that have been fused to corresponding affinity ligands. In this paper, we highlight the use of Src homology 3 (SH3) domains and corresponding peptide ligands in EMAC that have differential binding affinities towards SH3 for efficient capture and elution of proteins. Furthermore, differences between capture and elution of a monomeric and a multimeric protein were also studied.
authors.affiliation: Chemical and Biomolecular Engineering Department, University of Delaware, 150 Academy St., Newark, DE 19716, United States.
authors.forename: Heejae
authors.initials: H
authors.lastname: Kim
authors.affiliation: Chemical and Biomolecular Engineering Department, University of Delaware, 150 Academy St., Newark, DE 19716, United States. Electronic address: wilfred@udel.edu.
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D004549:Elastin / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D008024:Ligands; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000302:isolation & purification; D018909:src Homology Domains / Q000235:genetics*
chemical_list: D008024:Ligands; D010455:Peptides; D011994:Recombinant Proteins; D004549:Elastin
keywords: Affinity based purification; Elastin-like polypeptides; Non-chromatographic purification; Protein purification; SH3 domain