重组人促卵泡激素体外生物学活性测定方法的联合验证  

Collaborative study to evaluate a reporter gene assay for recombinant human follicle-stimulating hormone bioactivity

作　　者：王绿音 吕萍[1] 张慧[1] 李晶[1] 梁成罡[1] WANG Lü-yin;LU Ping;ZHANG Hui;LI Jing;LIANG Cheng-gang(Division of Hormone,National Institutes for Food and Drug Control,NHC Key Laboratory of Research on Quality and Standardization of Biotech Products,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院激素室,国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室,北京102629

出　　处：《药学学报》2023年第3期760-766,共7页Acta Pharmaceutica Sinica

基　　金：2020年《中华人民共和国药典》药品标准提高课题(2020S005)。

摘　　要：为探索基于报告基因的重组人促卵泡激素体外生物学活性测定方法的标准化应用前景,进行实验室间的联合验证。按照《中华人民共和国药典》2020年版通则9401对该方法进行相对准确度、中间精密度、线性与范围等方法学验证;联合3个实验室采用本方法测定来自不同生产企业的6批制剂及3批原液的体外生物学活性,对测定结果进行实验室间一致性分析以及实验室内、实验室间精密度分析;据联合验证中发现的问题对方法进行调整优化。实验室内方法学验证的结果符合《中华人民共和国药典》2020版四部通则9401的要求;针对联合验证中发现的问题,对细胞铺板培养基、标准品及供试品的预稀释缓冲液、加药后吸弃上清方式进行了优化,优化后各实验室之间的测定结果无显著差别(P>0.05),具有统计学等效性;实验室内及实验室间精密度良好,几何变异系数(GCV%)均小于15%。综上,本方法具有良好的实验室内重复性及实验室间重现性,且适用于不同生产厂家的重组人促卵泡激素制剂与原液产品,有望作为测定此类产品体外生物学活性的标准化方法进行推广。The goal of this work was to explore the prospect of standardized application of an in-vitro bioactivity assay for recombinant human follicle-stimulating hormone based on a reporter gene.The relative accuracy,intermediate precision,linearity and applicable range of the method were validated according to the General Rules of Chinese Pharmacopoeia 2020 edition Volume IV(9401).Three laboratories used this method to determine the in-vitro biological activities of six batches of drug product and three batches of drug substance manufactured by two different companies.The consistency of the potency determined by three laboratories,the intra-laboratory precision and inter-laboratory precision were analyzed.The method was optimized during the collaborative validation.The results of method validation meet the requirements of the General Rules of Chinese Pharmacopoeia 2020 edition Volume IV(9401).Aiming to resolve the problems found in the collaborative validation,the medium for cell seeding,the pre-diluted buffer solution of standard and sample,and the means of removing and discarding supernatant after stimulation were optimized.After optimization,there was no significant difference in the bioactivity among the different laboratories(P>0.05),indicating statistical equivalency.Intralaboratory and inter-laboratory precision were good and the geometric coefficient of variation(GCV%)were both less than 15%.In conclusion,the reporter gene assay has good intra-laboratory repeatability and inter-laboratory reproducibility and is suitable for analyzing recombinant human follicle-stimulating hormone drug product and drug substance by different manufacturers.It is expected to be used as a standardized method for the determination of the in-vitro bioactivity of such products.

关 键 词：生物学活性 重组人促卵泡激素 报告基因法 方法学验证 联合验证 

分 类 号：R917[医药卫生—药物分析学]