重组双功能水蛭素质量标准研究

以生色底物法测定抗凝血酶活性,比浊法测定抗血小板聚集活性,还原型SDS-PAGE测定分子量,SDS-PAGE和反相高效液相色谱测定纯度,毛细管电泳法测定等电点,胰蛋白酶酶切后进行肽图分析,其余检测项目按《中国药典》2005版三部规定进行。结果显示,用建立的方法对原液和成品进行了检定,各项指标均符合《人用重组DNA制品质量控制技术指导原则》和《中国药典》2005版三部的要求。建立的质控方法和质量标准具有保证产品安全、有效、质量可控的特点,可用于重组双功能水蛭素产品的常规检定。

Study on requirements for quality control of RGD-Hirudin

Anti-thrombin activity of RGD-Hirudin was determined by TH chromogenic substrate assay.Anti-platelet aggregation activity was determined by turbidimetry assay.Molecular weight of RGD-Hirudin was detected by reduced Tricine-SDS-PAGE.Purity of RGD-Hirudin was analysed by Tricine-SDS-PAGE and RP-HPLC.Isoelectic point and peptide mapping were detected by CE and tryptic digestion individually.Routine tests were all analysed according to Chinese Pharmacopoiea.The requirements and methods for quality control of RGD-Hirudin had been established and used for both RGD-Hirudin bulk and final product.The results of all tests complied with Chinese Pharmacopoiea and Guidelines for Quality Control of Recombinant DNA Products.The methods and requirements for quality control of RGD-Hirudin could assure the products safety and efficiency,and had been used for routine quality control of RGDHirudin.