Evaluation of chromatographic recycling for imidazole used in the chromatographic purification of His-tag recombinant proteins

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
A M NoubhaniX Santarelli

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.

References

Aug 1, 1992·Protein Expression and Purification·J Porath
Sep 1, 1983·Archives of Biochemistry and Biophysics·J PorathB Granstrand
Oct 23, 1997·Nature Biotechnology·T Misteli, D L Spector
Oct 6, 1998·Annual Review of Biochemistry·R Y Tsien
Feb 19, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·S GibertX Santarelli
Apr 8, 1960·Biochimica Et Biophysica Acta·J PORATH

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Citations

Sep 7, 2006·Journal of the American Chemical Society·Byung-Keun OhChad A Mirkin
Mar 30, 2006·Angewandte Chemie·Sarah J HurstChad A Mirkin
Feb 6, 2007·Colloids and Surfaces. B, Biointerfaces·Sylvia DaunertSapna Deo

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