促红细胞生成素糖基化的等电聚焦电泳分析  

Analysis of glycosylation of erythropoietin by isoelectric focusing electrophoresis

作　　者：潘婷琪[1] 易小萍[1] 孙祥明[1] 张元兴[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《中国生物制品学杂志》2012年第1期115-117,122,共4页Chinese Journal of Biologicals

摘　　要：目的建立一种用于促红细胞生成素(Erythropoietin,EPO)糖基化分析的更灵敏、更稳定的检测方法。方法比较固相pH梯度与载体两性电解质pH梯度等电聚焦电泳在灵敏度及EPO糖基化分析上的差异,并考察固相pH梯度等电聚焦电泳条带分布与唾液酸含量的关系。结果固相pH梯度等电聚焦电泳的灵敏度和分辨率明显高于载体两性电解质pH梯度等电聚焦电泳,可以更为客观地反映EPO的糖基化不均一性。固相pH梯度等电聚焦电泳的条带分布与唾液酸含量的测定结果基本一致。结论固相pH梯度等电聚焦电泳是一种更为理想的糖基化分析方法,可以有效分析EPO及其他重组蛋白药物的糖基化程度。Objective To develop a more stable and more sensitive method for analysis of glycosylation of erythropoietin （EPO）. Methods Immobilized pH gradient isoelectric focusing electrophoresis（IEF） and carrier ampholyte IEF were compared for sensitivity and analysis results of glycosylation of EPO, based on which the relationship between distribution of bands on immobilized pH gradient IEF profile and sialic acid content was investigated. Results Both the sensitivity and resolution of immobilized pH gradient IEF were significantly higher than those of carrier ampholyte pH gradient IEF, which reflected the heterogeneity of EPO glycosylation more objectively. The distribution of bands on immobilized pH gradient IEF profile was basically consistent with the determination result of sialic acid content. Conclusion Immobilized pH gradient IEF was a more ideal method for effective analysis of glycosylation of EPO and other recombinant proteins.

关 键 词：固相pH梯度等电聚焦电泳 载体两性电解质pH梯度等电聚焦电泳 促红细胞生成素 糖基化 

分 类 号：R392-33[医药卫生—免疫学]