purification and analysis of carbohydrate cluster from erythropoietin recombinant expression result on the leaven system of pichia pastoris

For clinical purposes, pure protein and identification of carbohydrate structure from erythropoietin recombinant are needed. Purification was done with His-Trap affinity chromatography method and continued with gel filtration chromatography column to get purer protein. The carbohydrate cluster from the resulting pure protein then can be recognized by using N- and O-glycosidase and can be compared to EPO recombinant from mammal cells. The result showed similarity on the declining trend of the protein molecule’s weight, which could be seen using the Western blot method. Pure oligosaccharide was hydrolyzed to produce various monosaccharide through incubation with HCl 4 N in 100 oC temperature for 6 hours and the result was applied on high intensity liquid chromatography incubator to learn the composition of its monosaccharide.