A single-step chromatographic method for the purification of proteins devoid of exposed histidine residues.
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.
Sharma
Surbhi
S
Downstream Processing Lab, Institute of Genomics and Integrative Biology, Delhi, India.
Gade
W N
WN
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*
D007098:Imino Acids; D008670:Metals; D006639:Histidine; D007986:Luteinizing Hormone; C008109:iminodiacetic acid