Immobilized metal-ion affinity chromatography of human growth hormone

✍ Scribed by Federico Maisano; Silvia Astrua Testori; Guido Grandi

• Publisher: Elsevier Science
• Year: 1989
• Tongue: English
• Weight: 318 KB
• Volume: 472
• Category: Article
• ISSN: 1873-3778
• DOI: 10.1016/s0021-9673(00)94144-x

✦ Synopsis

Affinity chromatography on immobilized metal ions (IMAC) is a separation technique that takes advantage of the different capabilities of proteins of interacting with metal ions mainly through their exposed histidine residues. The metal ions are immobilized on suitable supports through a chelating ligand, usually iminodiacetate, which is covalently bound to the matrix . This technique was introduced by Porath et al.' in 1975 and since then it has been adopted for the purification of many proteins . Recent advances with this method include the semipreparative purification of proteins from cell culture filtrates' and high-performance applications' .

We have studied the behaviour of the recombinant human growth hormone (hGH) when subjected to IMAC and found this technique to be particularly useful for hGH purification . Further, by comparing our results with those observed for some model proteins', we have attempted to draw some conclusions about the number and location of histidine residues exposed on the molecule surface .