酶联免疫法测定甘精胰岛素中单链前体残留研究  

Detection of single-chain precursor residues in insulin glargine by ELISA

作　　者：张孝明 吕萍 胡馨月 丁晓丽 李晶 梁成罡 ZHANG Xiaoming;LYU Ping;HU Xinyue;DING Xiaoli;LI Jing;LIANG Chenggang(Institute for Chemical Drug Control,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化学药品检定所,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：《中国药物警戒》2024年第3期290-294,共5页Chinese Journal of Pharmacovigilance

基　　金：国家重点研发计划(2021YFF0600804)。

摘　　要：目的 建立酶联免疫法检测甘精胰岛素中单链前体残留,为甘精胰岛素药物质量控制和安全性评价提供新方法。方法采用双抗夹心酶联免疫法,以甘精胰岛素原为标准品,建立单链前体残留检测方法,验证方法的重复性、准确度、范围和精密度,并应用该方法检测9批甘精胰岛素原料药。结果在0.156~10.00 ng·mL^(-1)浓度范围内,对甘精胰岛素原标准品进行四参数曲线拟合,曲线相关系数R~2大于0.99,方法准确度较高、重复性良好。重复检测3批甘精胰岛素原料药,平均加标回收率在104%~113%范围内,RSD%小于10%,方法精密度良好。9批样品中单链前体残留量均小于1 ng·mg^(-1)。结论建立的酶联免疫法可用于甘精胰岛素及其他重组人胰岛素类似物中单链前体残留量的质量控制和安全性评价。Objective To establish an enzyme-linked immunoassay for the detection of single-chain precursor residues in insulin glargine so as to recommend a new method for quality control and safety evaluation of insulin glargine.Methods An enzyme-linked immunoassay based on double antibody sandwich was established by using proinsulin glargine as the standard.The repeatability,accuracy,scope and precision of the method were verified,and nine batches of insulin glargine drug substances were tested using this method.Results Four-parameter curve fitting was performed using the proinsulin glargine standard at concentrations ranging from 0.156 to 10.00 ng·mL^(-1),and the correlation coefficient of the curve exceeded 0.99.This method was highly accurate and repeatable.Three batches of insulin glargine drug substances were repeatedly detected.The average recovery ranged from 104%to 113%,and the RSD%was less than 10%,suggesting the good precision of this method.The single-chain precursor residues in all the nine batches were less than 1 ng·mg^(-1).Conclusion The enzyme-linked immunoassay established in this study can be used for quality control and safety evaluation of single-chain precursor residues in insulin glargine and other recombinant human insulin analogues.

关 键 词：酶联免疫法 甘精胰岛素 单链前体 C肽 质量控制 方法学验证 安全性 回收率 

分 类 号：R917[医药卫生—药物分析学]