Abstract

For clinical purposes, pure protein and identification of carbohydrate structure from recombinant erythropoietin are needed. Purification was done by Immobilized Metal Affinity Chromatography (IMAC) column charged with Ni2+ (His-Trap affinity chromatography) and continued with gel filtration chromatography column to get purer protein. The carbohydrate group which is oligosaccharide from the resulting pure protein then can be recognized by using N- and O-glycosidase. Pure oligosaccharide was hydrolyzed to produce various monosaccharide through incubation with 4 N HCl in 100 oC temperature for 6 hours and the result was applied on High Performance Liquid Chromatography (HPLC) column to learn the composition of its monosaccharide. Keywords: EPO, gel filtration chromatography, His-Trap affinity chromatography, monosaccharide

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