论文部分内容阅读
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 a quality control method and quality standard for recombinant human granulocyte colony-stimulating factor-Fc fusion protein. Methods: The biological activity was determined by NSF-60 cell proliferation assay. Purity was determined by non-reducing SDS-PAGE and RP-HPLC. Residual CHO cell protein and protein A were determined by ELISA. The remaining test items were according to the 2010 edition of Chinese Pharmacopoeia Regulations. Results: The recombinant human granulocyte colony-stimulating factor-Fc fusion protein and the finished product were assayed by the established method. All the indexes were in line with the “Guiding Principles for Quality Control of Human Recombinant DNA Products” and the 2010 edition of Chinese Pharmacopoeia Claim. Conclusion: The established quality control methods and quality standards have the characteristics of ensuring the safety and effectiveness of the products and controlling the quality of the products, and can be used for the routine verification of such products.