聚乙二醇重组人血管内皮抑制素质控方法的建立

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8...

Development of A Method for Quality Control of PEGylated Recombinant Human Endostatin

Objective To develop a method for quality control of PEGylated recombinant human endostatin(PEG-ES).Methods Test samples were determined for biological activity by fluorimetric endothelial cell migration assay and for purity and content by RP-HPLC.Peptide mapping was carried out by trypsin digestion,and other tests according to the requirements in Chinese Pharmacopoeia(Volumes Ⅱ and Ⅲ,2005 edition).Results The specific activities of three batches of bulks of PEG-ES determined by the developed method were 88.4,56.5 and 89.6 U / mg,while the activities of final products were 81%,92% and 151% of stated amount,respectively.The protein contents of three batches of final products determined by RP-HPLC were 101.0%,97.0% and 98.1% of stated amount respectively.All the purities of three batches of bulks determined by RP-HPLC and SDS-PAGE were more than 99.9%.All the results of peptide mapping were consistent with those of control,and all the other quality indexes met the requirements in Chinese Pharmacopoeia.Conclusion The developed method laid a foundation of effective quality control of PEG-ES.