重组人激肽释放酶-1质量标准研究  

Study on methods and requirements for quality control of recombinant human kallikrein-1

作　　者：李响[1] 韩春梅[1] 李永红[1] 毕华[1] 高凯[1] 丁有学[1] 饶春明[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《药物分析杂志》2010年第9期1676-1679,共4页Chinese Journal of Pharmaceutical Analysis

基　　金：国家"863"高技术发展计划项目(课题编号2003AA2Z3481)

摘　　要：目的:建立重组人激肽释放酶-1(rhK1)的质控方法和质量标准。方法:以S2266底物显色法测定rhK1的酶活性,还原型SDS-PAGE测定相对分子质量,非还原型SDS-PAGE测定相关蛋白,SDS-PAGE和反相高效液相色谱法测定纯度和成品含量,平板水平等电聚焦电泳测定等电点,用胰蛋白酶酶切后分析肽图,其余检测项目按2005年版中国药典三部规定进行。结果: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2005年版中国药典三部的要求。结论: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rhK1产品的常规检定。Objective：To establish methods and requirements for quality control of recombinant human kallikrein-1（rhK 1）.Methods：Enzyme activity potency of rhK 1 was determined by substance S2266 development assay;Relative molecular mass of rhK 1 was detected by reduced SDS-PAGE;Content of related protein was tested by non-reduced SDS-PAGE;Purity and content of final product were analyzed by SDS-PAGE and RP-HPLC;Isoelectic point and peptide mapping were detected by IEF and tryptic digestion individually.Routine test were all analysed according to the Chinese Pharmacopoiea（2005）.Results：The requirements and methods for quality control of rhK 1 had been established and used for both rhK 1 bulk and final product.The results of all test complied with Chinese Pharmacopoiea and Guidelines for Quality Control of Recombinant DNA Products.Conclusion：The methods and requirements for quality control of rhK 1 can assure the products safety and efficiency,and has been used for routine quality control of rhK 1.

关 键 词：重组人激肽释放酶-1(rhK1) 质量控制 

分 类 号：R917[医药卫生—药物分析学]