title: A single-step chromatographic method for the purification of proteins devoid of exposed histidine residues.
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.
authors.affiliation: Downstream Processing Lab, Institute of Genomics and Integrative Biology, Delhi, India.
authors.forename: Surbhi
authors.initials: S
authors.lastname: Sharma
authors.forename: W N
authors.initials: WN
authors.lastname: Gade
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006639:Histidine / Q000737:chemistry*; D007098:Imino Acids / Q000737:chemistry*; D007986:Luteinizing Hormone / Q000302:isolation & purification*; D008670:Metals / Q000737:chemistry*; D010902:Pituitary Gland / Q000737:chemistry*
chemical_list: D007098:Imino Acids; D008670:Metals; D006639:Histidine; D007986:Luteinizing Hormone; C008109:iminodiacetic acid