Purification of recombinant proteins with metal chelate adsorbent.

In 1975 Porath and co-workers introduced immobilized metal ion affinity chromatography (IMAC) for the purification of peptides and proteins (1). The principle of this technique is the coordination between the electron donor groups on a protein (peptide) surface and immobilized transition metal ions. The tridentate chelator, iminodiacetic acid, is coupled via a spacer arm to a solid support and used for the immobilization of metal ions such as Ni(II), Cu(II) or Zn(II). Porath postulated that the histidine, cysteine and tryptophan residues in proteins (peptides) are most likely to form stable coordination bonds with metal chelates at neutral pH. Present experience (2) indicates that histidine residues on protein surfaces are the predominant electron donor groups.

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