Purifikasi dan Pencirian Eritropoietin Rekombinan Hasil Ekspresi pada Sistem Khamir Pichia pastoris

Abstract

For the clinical purposes needed a purified protein and the carbohydrates contains of recombinant erythropoietin (EPO) should be known. Purification was done by histrap method and then applied to column chromatography gel filtration to make more purified. Carbohydrates groups which is oligosaccharide on purified protein then was characterized by N- and O- glycosidase and the pattern compared with recombinant EPO by CHO cells. The result show has a similar molecular mass which visualized by Western blot method. The purified oligosaccharide were hydrolysed to monosaccharides by incubation in 4 N HCl at 100 ºC for 6 h in evacuated sealed tube. Monosaccharides were separated by high performance liquid chromatography instrument, but unfortunately, the results cannot be shown the kind of monosaccharides.