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目的建立聚乙二醇重组人血管内皮抑制素的质控方法。方法采用内皮细胞迁移荧光分析法测定样品的生物学活性;反相高效液相色谱(RP-HPLC)法测定样品的纯度和含量;胰酶消化法测定肽图;其余项目按《中国药典》三部(2005版)和二部进行检测。结果用建立的生物学活性测定方法测定的3批原液的比活性分别为88.4、56.5和89.6 U/mg,3批成品的活性分别为标示量的81%、92%和151%;RP-HPLC法测定的3批成品蛋白含量分别为标示量的101.0%、97.0%和98.1%;RP-HPLC法和SDS-PAGE法测定的3批原液的纯度均大于99.9%;肽图分析结果均与对照品一致;其余各项指标均符合规定。结论所建立的质控方法为有效地控制聚乙二醇重组人血管内皮抑制素的质量奠定了基础。
Objective To establish a quality control method for recombinant human vascular endostatin. Methods The biological activity of the sample was determined by fluorescence assay of endothelial cell migration. The purity and content of the sample were determined by reverse-phase high-performance liquid chromatography (RP-HPLC). The peptide map was determined by trypsin digestion. Department (2005 Edition) and two for testing. Results The specific activity of the three batches of crude solutions determined by the established biological activity assay were 88.4, 56.5 and 89.6 U / mg, respectively. The activity of the three batches of products was 81%, 92% and 151% of the labeled amount, respectively. The three batches of crude protein were 101.0%, 97.0% and 98.1% of the labeled amount, respectively. The purity of the three batches of crude extracts measured by RP-HPLC and SDS-PAGE were more than 99.9% Consistent products; the rest of the indicators are in line with the provisions. Conclusion The established quality control method lays the foundation for effectively controlling the quality of recombinant human endostatin.