title: Purification and on-column refolding of EGFP overexpressed as inclusion bodies in Escherichia coli with expanded bed anion exchange chromatography.
abstract: The enhanced green fluorescent protein (EGFP) was over-expressed in Escherichia coli as inclusion bodies to increase its quantity and to facilitate its purification. Insoluble EGFP has been purified on Q Hyper Z matrix by expanded bed adsorption after solubilization in 8 M urea. The adsorption was made in expanded bed mode to avoid centrifugation. EBA-column refolding was done by elimination of urea and elution with NaCl. The EGFP was obtained as a highly purified soluble form with similar behavior in fluorescence and electrophoresis as native EGFP.
authors.affiliation: Ecole Supérieure de Technologie des Biomolécules de Bordeaux (ESTBB), Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France. charlotte.cabanne@estbb.u-bordeaux.fr
authors.forename: C
authors.initials: C
authors.lastname: Cabanne
authors.forename: A M
authors.initials: AM
authors.lastname: Noubhani
authors.forename: A
authors.initials: A
authors.lastname: Hocquellet
authors.forename: F
authors.initials: F
authors.lastname: Dole
authors.forename: W
authors.initials: W
authors.lastname: Dieryck
authors.forename: X
authors.initials: X
authors.lastname: Santarelli
mesh_terms: D000327:Adsorption; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism* / Q000648:ultrastructure; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification*; D002479:Inclusion Bodies / Q000378:metabolism; D017510:Protein Folding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012995:Solubility; D013050:Spectrometry, Fluorescence; D014508:Urea
chemical_list: D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D014508:Urea