Evaluation of chromatographic recycling for imidazole used in the chromatographic purification of His-tag recombinant proteins.
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.
Noubhani
A M
AM
Ecole Supérieure de Technologie des Biomolécules de Bordeaux (ESTBB), UMR 5544, Université Victor Segalen Bordeaux 2, 146 Rue Léo Saignat, 33076 Cedex, Bordeaux, France.
Dieryck
W
W
Bakalara
N
N
Latxague
L
L
Santarelli
X
X
D002846:Chromatography, Affinity / Q000379:methods*; D006639:Histidine / Q000737:chemistry*; D007093:Imidazoles / Q000032:analysis*; D011994:Recombinant Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results
D007093:Imidazoles; D011994:Recombinant Proteins; D006639:Histidine