PURIFICATION AND CARBOHYDRATE ANALYSIS OF RECOMBINANT HUMAN ERYTHROPOIETIN EXPRESSED IN YEAST SYSTEM Pichia pastoris

被引：2

作者：

Wardiana, Andri ^{[1
]}

Santoso, Adi ^{[1
]}

机构：

[1] Indonesian Inst Sci LIPI, Biotechnol Res Ctr, Cibinong 16911, Jawa Barat, Indonesia

来源：

MAKARA JOURNAL OF SCIENCE | 2011年 / 15卷 / 01期

关键词：

EPO; gel filtration chromatography; His-Trap affinity chromatography; monosaccharide;

D O I：

10.7454/mss.v15i1.888

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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 Oglycosidase. Pure oligosaccharide was hydrolyzed to produce various monosaccharide through incubation with 4 N HCl in 100 degrees C temperature for 6 hours and the result was applied on High Performance Liquid Chromatography (HPLC) column to learn the composition of its monosaccharide.

引用

页码：75 / 78

页数：4