重组人促红细胞生成素聚乙二醇修饰及纯化研究

Modification of Recombinant Human Erythro-protein(EPO) with Polyethylene Glycol(PEG) and Purification of PEG-EPO

目的 研究重组人促红细胞生成素 (rhEPO)聚乙二醇修饰条件及纯化方法。方法 用SDS PAGE分析不同反应条件对产物成分的影响 ;用红细胞压积法测定各组分的体内生物学活性 ;用分子筛柱层析进行纯化。结果 rhEPO的修饰度随蛋白浓度、蛋白与PEG2 NHS活性酯质量比的增加而提高 ,体内生物学活性随修饰度增加而降低 ,分子筛柱层析纯化效果好。

Objective To explore the condition for the modification of recombinant human erythro poietin (EPO) with polyethylene glycot(PEG) and the method for the purification of PEG EPO.Methods Analyze the influence of various conditions on the components of PEG EPO by SDS PAGE,detect the bioactivity in vivo of PEG EPO by hematocrit method and purify PEG EPO by molecular sieve column chromatography.Results The modification degree of rhEPO increased with the increasing protein concentration as well as the increasing mass ratio of protein to PEG 2 NHS active ester.However,the bioactivity in vivo of PEG EPO decreased with the increasing modification degree.Molecular sieve column showed good purification effect.Conclusion The factors influencing the modification of EPO with PEG were determined,and a purification method for PEG EPO was developed.