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抗白介素-1β单克隆抗体生物学活性检测方法的建立
抗白介素-1β单克隆抗体生物学活性检测方法的建立
刘春雨,徐刚领,于传飞,张 峰,王文波,李 萌,陈 伟,王 兰,高 凯

中国食品药品检定研究院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
Development of a method for determination of potency of anti-IL-1β monoclonal antibody
(National Institutes for Food and Drug Control, Beijing 100050, China)

摘要参考文献相关文章

起始页:1959

摘要:目的:建立抗白介素-1β单克隆抗体(抗IL-1β单抗)的生物学活性检测方法。方法:利用HEK293-NF-κb-luc细胞系,通过荧光素酶检测系统(Steady-Glo Luciferase Assay System)进行抗IL-1β单抗的生物学活性检测,以抗IL-1β单抗参比品通过平行线分析的方法计算供试品相对效价,并对该方法进行精密性和准确性验证。结果:抗IL-1β单抗供试品及参比品该方法中均存在量效关系,在半对数坐标纸上呈平行的直线分布。5批抗IL-1β单抗成品经3 次测定,相对效价平均值在(84.57±3.46)%~(101.93±7.56)%之间,变异系数均小于10%。2批回收率样品经3 次测定,回收率分别为(113.00±15.32)%和(108.53±23.51)%。结论:已成功建立抗IL-1β单抗生物学活性检测方法,该方法重复性好,准确性高,可作为抗IL-1β单抗生物学活性的常规检测方法。

关键词:抗IL-1β单克隆抗体;生物学活性;荧光素酶检测系统

通讯作者:王兰,女,博士,副研究员,主要从事抗体类生物治疗药物质量控制研究。

基金项目:国家“重大新药创制”科技重大专项(2014ZX09304311-001,2012ZX09304010)

作者简介:刘春雨,女,硕士,助理研究员,主要从事抗体类生物治疗药物质量控制研究。

Abstract:Objective:To develop a method for determination of biological activity of anti-IL-1β monoclonal antibody. Methods:The biological activity of anti-IL-1β mAb was determined by Steady-Glo Luciferase Assay System using HEK293-NF-κb-luc cell line, and the relative potency was calculated by comparing with the reference using the method of parallel analysis. The developed method was verified for precision and accuracy. Results:Either drug prodct or reference of anti-IL-1β mAb showed a dose-response mode by the developed method, presenting parallel straight lines on semi-log coordinate paper. Five batches of f drug product of anti-IL-1β mAb were determined for 3 times, and the result showed mean relative potencies of (84.57±3.46)%~(101.93±7.56)%, with a variation coefficient of less than 10%. Two batches of drug product were determined for 3 times, and the result showed recovery rates of (113.00±15.32)% and (108.53±23.51)% respectively. Conclusion:A method for determination of biological activity of anti-IL-1β mAb was successfully developed, which showed good reproducibility and high accuracy and might be used routinely.

Key words:anti-IL-1β monoclonal antibody; biological activity; Steady-Glo Luciferase Assay System



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