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目的对我国已上市的重组人促红细胞生成素的一级结构进行比对分析。方法针对9家制药企业生产的重组人促红细胞生成素理化对照品,测定质谱相对分子质量、液质肽图、寡糖图谱及唾液酸含量,并对测定结果进行比对分析。结果各样品去除N-糖基后实测相对分子质量均与理论值一致,误差均小于1;液质肽图测定中氨基酸覆盖率均为100%,序列均与理论一致;二硫键连接方式均为Cys7-Cys161、Cys29-Cys33;O-糖基化位点均为Ser126,N-糖基化位点均为Asn24、Asn38、Asn83;各样品的糖基修饰方式存在差异,不同糖型的相对比例不一致;唾液酸含量测定结果介于(物质的量的比11.5~15.7)促红细胞生成素之间。结论 9家EPO制品的一级结构基本一致,差异主要表现在糖基修饰水平。
Objective To compare the primary structure of recombinant human erythropoietin in China. Methods The recombinant human erythropoietin reference substance produced by nine pharmaceutical companies was determined. The relative molecular mass, liquid peptide map, oligosaccharide profile and sialic acid content of the recombinant human erythropoietin were determined. The results were compared and analyzed. Results The measured relative molecular mass of all samples after removing N-glycosyl were consistent with the theoretical values, the errors were less than 1; the amino acid coverage in liquid peptide map was 100%, the sequences were consistent with the theory; disulfide bond Were Cys7-Cys161, Cys29-Cys33; O-glycosylation sites were Ser126, N-glycosylation sites were Asn24, Asn38, Asn83; Glycosylation of each sample there are differences in the way, the different glycoforms relative The proportion of inconsistent; sialic acid content determination results (substance ratio of 11.5 to 15.7) between erythropoietin. Conclusions The primary structures of nine EPO products are basically the same, the differences are mainly manifested in the level of glycosylation.