重组人脑利钠肽质量标准与检定方法研究

Study of the Requirements and Methods for Quality Control of Recombinant Human Brain Natriuretic Peptide

目的:建立重组人脑利钠肽(rhBNP)质量标准与检定方法,用于该产品的质量控制。方法:用离体动脉条测定法测定rhBNP的生物活性;用反相HPLC和SDS-PAGE分析蛋白纯度;用肽图、免疫印迹及N-端氨基酸序列测定分析、鉴定产品特性;用固相斑点杂交法测定残余外源DNA含量;用ELISA测定残余工程菌蛋白含量;用《中国生物制品规程》的方法对,rhBNP成品进行无菌、热原、细菌内毒素和异常毒性等常规试验等。结果:用建立的方法对连续3批rhBNP原液和成品进行检定,结果各项指标均符合《重组DNA制品质量控制要点》和《中国生物制品规程》的要求。结论:建立的rhBNP质量标准与检定方法,具有保证产品安全、有效、质量可控等特点,可用于该产品的常规检定。

Objective: To establish the requirements and methods for quality control of Recombinant Human Brain Natriuretic Peptide (rhBNP). Methods: Biological activity was determined by tension change of artery in vitro. The purity of protein was analysed by RP - HPLC and SDS - PAGE. Identification tests were done by peptide mapping, western blot and N - terminal amino acid sequence. Residual DNA and residual proteins of host cell were separately detected by DNA hybridization and ELISA. Routine tests like sterility test, pyrogen test, bacterial en-dotoxin test and abnormal toxicity test were all fulfilled accordingly to the National Requirements for Biological Products. Results: The requirements and methods for quality control of rhBNP have been established and used for quality control of three simultaneous batches of bulk and final product rhBNP. The results of all tests complied with the requirements of quality control of rhBNP. Conclusion: The requirements and methods for quality control of rhBNP showed the advantages of assuring the product safety and efficiency, which can be used for routine quality control of rhBNP.