title: Evaluation of chromatographic recycling for imidazole used in the chromatographic purification of His-tag recombinant proteins.
abstract: The aim of this work was to test a recycling method for imidazole used in immobilized metal affinity chromatography (IMAC) as eluent for recombinant histidine-tag (His-tag) protein. After evaluating two supports, the method was optimized with a mixture of bovine serum albumin, sodium chloride and imidazole. Recycling was performed with an eluate fraction from IMAC of His-tag enhanced green fluorescent protein produced in our laboratory and pure imidazole was recovered in water and was analyzed after being freeze-dried. The imidazole was then reused as eluent in IMAC without any modification in its structure or behavior. This procedure can be used for large-scale chromatography.
authors.affiliation: Ecole Supérieure de Technologie des Biomolécules de Bordeaux (ESTBB), UMR 5544, Université Victor Segalen Bordeaux 2, 146 Rue Léo Saignat, 33076 Cedex, Bordeaux, France.
authors.forename: A M
authors.initials: AM
authors.lastname: Noubhani
authors.forename: W
authors.initials: W
authors.lastname: Dieryck
authors.forename: N
authors.initials: N
authors.lastname: Bakalara
authors.forename: L
authors.initials: L
authors.lastname: Latxague
authors.forename: X
authors.initials: X
authors.lastname: Santarelli
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D006639:Histidine / Q000737:chemistry*; D007093:Imidazoles / Q000032:analysis*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results
chemical_list: D007093:Imidazoles; D011994:Recombinant Proteins; D006639:Histidine