重组人粒细胞集落刺激因子-Fc融合蛋白(rhG-CSF-Fc)的质量研究  

Quality control research of recombinant human granulocyte colony-stimulating factor-Fc fusion protein(rhG-CSF-Fc)

作　　者：高凯[1] 王兰[1] 陶磊[1] 于雷[1] 李响[1] 史新昌[1] 刘兰[1] 饶春明[1] 

机构地区：[1]中国食品药品检定研究院重组技术产品室,卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《药物分析杂志》2012年第1期107-110,共4页Chinese Journal of Pharmaceutical Analysis

基　　金：国家“重大新药创制”科技重大专项(2009ZX09307-001)

摘　　要：目的:建立重组人粒细胞集落刺激因子-Fc融合蛋白的质控方法和质量标准。方法:采用NSF-60细胞增殖法测定生物学活性;非还原SDS-PAGE电泳和RP-HPLC测定纯度;以ELISA法分别测定残留CHO细胞蛋白和蛋白A;其余检测项目按中国药典2010年版三部规定进行。结果:用建立的方法对重组人粒细胞集落刺激因子-Fc融合蛋白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2010年版三部的要求。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,可用于该类产品的常规检定。Objective：To establish methods and requirements for quality control of recombinant human granulocyte colony - stimulating factor - Fc fusion protein （ rhG - CSF - Fc ）. Methods： Biological activity of rhG - CSF - Fc was determined by NFS -60 cell proliferation assay. Non reducing SDS - PAGE and RP - HPLC were applied to analyze the product purity. CHO host cell protein and Protein A residue were determined by ELISA. Routine tests were all carried out according to Pharmacopoeia of People＇s Republic of China（ volume Ⅲ ,2010 edition）. Results： The requirements and methods for quality control of rhG - CSF - Fc were established and used for quality control of bulk and final product. The results of all tests complied with the requirements of Guideline for Quality Control of Recombinant DNA Products for Human Use and Pharmacopoeia of People＇ s Republic of China （ volume Ⅲ ,2010 edition）. Conclusion：The methods and requirements for quality control of rhG - CSF - Fc showed characteristics of assuring the products safety and efficacy,which can be used for routine quality control rhG -CSF -Fc.

关 键 词：重组人粒细胞集落刺激因子-Fc融合蛋白 十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE) 反相高效液相色谱 酶标记免疫吸附测定(ELISA) 等电聚焦电泳 肽图分析 质量控制 生物学活性 

分 类 号：R917[医药卫生—药物分析学]