重组人肿瘤坏死因子相关凋亡诱导配体(rhTRAIL)质控方法和质量标准的研究  

Study on Methods and Requirements for Quality Control of Recombinant Human Tumor Necrosis Factor Related Apoptosis Inducing Ligand(rhTRAIL)

作　　者：高凯[1] 杨婉娟[2] 韩春梅[2] 丁有学[2] 饶春明[2] 张英起[1] 王军志[1] 

机构地区：[1]中国人民解放军第四军医大学研究生院,西安730000 [2]中国药品生物制品检定所生化室,北京100050

出　　处：《药物分析杂志》2005年第3期253-257,共5页Chinese Journal of Pharmaceutical Analysis

基　　金：国家863高技术研究发展计划项目;课题编号:2003AA2Z3480

摘　　要：目的:建立重组人肿瘤坏死因子相关凋亡诱导配体(rhTRAIL)的质控方法和质量标准。方法:以NCI—H460细胞毒实验测定rhTRAIL的生物学活性,还原型SDS—PAGE确定相对分子质量,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,用胰蛋白酶酶切后分析肽图,其余检测项目按《中国生物制品规程》2000版规定进行。结果:用建立的方法对rhTRAIL原液和成品各1批进行了检定,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《中国生物制品规程》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,可用于重组人肿瘤坏死因子相关凋亡诱导配体(rhTRAIL)产品的常规检定。Objective:To establish methods and requirements for quality control of recombinant human tumor necrosis factor related apoptosis inducing ligand (rhTRAIL).Method:Biological potency of rhTRAIL was determined by NCI-H460 cell cytotoxity assay.relative molecular mass of rhTRAIL was detected by reducing SDS-PAGE.Purity of rhTRAIL was analyzed by SDS-PAGE and RP-HPLC.Isoelectric point and peptide mapping were detected separately by CE and tryptic digestion.Routine tests were all carried out accordingly to the National Requirements for Biological Products.Results:The requirements and methods for quality control of rhTRAIL had been established and used for quality control of bulk and final product.The results of all tests complied with the requirements of quality control of rhTRAIL.Conclusion:The methods and requirements for quality control of rhTRAIL showed advantages of assuring the products safety and efficacy,which can be used for routine quality control of rhTRAIL.

关 键 词：TRAIL 质控方法 相关凋亡诱导配体 重组人肿瘤坏死因子 NCI 生物制品 细胞毒 中国 产品安全 质量标准 

分 类 号：R730[医药卫生—肿瘤] R735.8[医药卫生—临床医学]