PMID: 11334341May 4, 2001Paper

Polymer versus monomer as displacer in immobilized metal affinity chromatography

Journal of Chromatography. B, Biomedical Sciences and Applications
P ArvidssonB Mattiasson

Abstract

Successful immobilized metal affinity chromatography (IMAC) of proteins on Cu2+-iminodiacetic acid Sepharose has been carried out in a displacement mode using a synthetic copolymer of vinyl imidazole and vinyl caprolactam [poly(VI-VCL)] as a displacer. Vinyl caprolactam renders the co-polymer with the thermosensitivity, e.g., property of the co-polymer to precipitate nearly quantitatively from aqueous solution on increase of the temperature to 48 degrees C. A thermostable lactate dehydrogenase from the thermophilic bacterium Bacillus stearothermophilus modified with a (His)6-tag [(His)6-LDH] has been purified using an IMAC column. For the first time it was clearly demonstrated that a polymeric displacer [poly(VI-VCL)] was more efficient compared to a monomeric displacer (imidazole) of the same chemical nature, probably due to the multipoint interaction of imidazole groups within the same macromolecule with one Cu2+ ion. Complete elution of bound (His)6-LDH has been achieved at 3.7 mM concentration of imidazole units of the co-polymer (5 mg/ml), while this concentration of free imidazole was sufficient to elute only weakly bound proteins. Complete elution of (His)6-LDH by the free imidazole was achieved only at concentrations as...Continue Reading

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Citations

Apr 2, 2002·Protein Expression and Purification·Sheryl FernandesRajni Hatti-Kaul
Feb 24, 2005·Chemical Society Reviews·Carolina de Las Heras AlarconCameron Alexander
Apr 6, 2018·Physical Chemistry Chemical Physics : PCCP·Reddicherla UmapathiPannuru Venkatesu
Mar 22, 2003·Journal of Chromatography. a·E K M UedaL Morganti

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