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Efficiently Isolate Your Protein


Metal Chelate Affinity Chromatography

Merck:/Freestyle/PS-Process-Solutions/BioP-Mfg-Biopharmaceutical-Manufacturing/PS-13-07972-226x159.jpg
Fig 1. Purification of glucokinase from yeast using Fractogel® EMD Chelate loaded with cobalt ions. Elution was achieved during a decreasing pH-gradient from pH 7.5 to pH 6.0 (buffer: 20 mM phosphate; 1 M KCI, 10 mM glucose).

Metal affinity chromatography is well established as an efficient means of purifying a variety of proteins and peptides. Special advantages of this pseudo-biospecific technique are the stability of the used ligands, and lower costs compared to most of the bio-ligands.

The technique is especially useful for isolation of proteins with exposed histidine, cysteine, and tryptophan residues, also making it useful for isolation of histidine-tagged proteins. Because the presence of high salt concentrations in the buffer does not interfere with binding performance, tentacle metal chelate chromatography is ideally used in combination with an ion exchanger column. The pooled fractions of an anion or cation exchanger column after a salt gradient elution can be applied directly to a Fractogel® EMD Chelate column for an efficient chromatography strategy.

Fractogel® Metal Chelate provides:

  • Efficient purification of His-tagged proteins
  • High protein binding capacity due to an optimal steric accessibility
  • Easy on-column refolding

Fractogel® EMD Chelate uses iminodiacetic acid as the metal chelating ligand. The iminodiacetic acid residue is very suitable as an immobilized chelating agent, since a bidentate chelating moiety remains free after immobilization, to which a metal ion can be bound. Various metal ions can be immobilized on the stationary phase via this immobilized chelating agent. Free binding sites of the metal ions are used to bind different proteins and peptides.



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