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  • Title: A single-step chromatographic method for the purification of proteins devoid of exposed histidine residues.
    Author: Sharma S, Gade WN.
    Journal: Prep Biochem Biotechnol; 2004 May; 34(2):145-52. PubMed ID: 15195709.
    Abstract:
    The principle of the immobilized metal affinity chromatography (IMAC) is based on the differences in the affinity of proteins for metal ions bound in a 1:1 complex of iminodiacetic acid (IDA) immobilized on a chromatographic support. A single step purification was carried out for luteinizing hormone (LH) on Cu2+, Zn2+, Ni2+, and Co2+ IDA Sepharose affinity columns. Highly purified LH was obtained with a Cu2+ IDA Sepharose column. SDS-PAGE and Western blot analysis were done to confirm the purity of the hormone. Biological activity has been evaluated by radio-immunoassay. This method was found economically viable and suitable for the recovery of biologically active hormone.
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