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 establish a more sensitive and stable method for the glycosylation analysis of erythropoietin (EPO). Methods The differences in sensitivity and EPO glycosylation between pH gradient and pH gradient isoelectric focusing electrophoresis of amphiphilic electrolytes were compared. The relationship between the bands distribution of solid phase pH gradient isoelectric focusing electrophoresis and sialic acid content was also investigated. Results The sensitivity and resolution of the solid-phase pH gradient isoelectric focusing electrophoresis were significantly higher than those of the carrier ampholyte pH gradient isoelectric focusing electrophoresis, which could more objectively reflect the glycosylation heterogeneity of EPO. The band distribution of solid-phase pH gradient isoelectric focusing electrophoresis and sialic acid content were basically the same. Conclusion The solid-phase pH gradient isoelectric focusing electrophoresis is a more ideal method for glycosylation analysis, which can effectively analyze the degree of glycosylation of EPO and other recombinant protein drugs.