重组人内皮抑素的质控方法研究

目的通过研究并建立重组人内皮抑素质量控制方法。方法用内皮细胞迁移法测定内皮抑素的生物学活性;胰酶消化法测定肽图;分别用非还原型SDS-PAGE和RP-HPLC法测定样品的纯度;其余项目按《中国生物制品规程》2000年版进行测定。结果建立了内皮抑素的活性测定方法,应用该方法测定３批内皮抑素比活性分别为1.45×106, 1.57×106和2.73×106 u·mg^-1蛋白,肽图测定结果显示3批样品批间一致,SDS-PAGE纯度和RP-HPLC纯度均大于99%,其余各项指标均符合规定。结论建立的方法可有效地控制内皮抑素制品的质量。

QUALITY CONTROL METHODS FOR RECOMBINANT HUMAN ENDOSTATIN

AIM: To establish the quality control methods for recombinant human endostatin. METHODS: Biological activity was determined by endothelial cell migration assays. Peptide mapping was tested by trypsin digestion and RP-HPLC. Purity was determined by non-reduced SDS-PAGE and RP-HPLC. Other tests including molecular weight, isoelectrical point, etc. were done according to the National Requirements for Biological Products (2000). RESULTS: The method of bioassay was established and used for determining activity of endostatin. Specific activity of the three batchs of drug substance was 1.45 x 10(6), 1.57 x 10(6) and 2.73 x 10(6) u.mg-1 proteins. Peptide mappings of the three batches of drug substance were completely identified. Both purity results of the products tested by SDS-PAGE and RP-HPLC were more than 99%. CONCLUSION: The established methods can effectively control the quality of recombinant human endostatin.