微量动态显色法(mKC)检测抗体药物中细菌内毒素含量方法验证及对比实验

Verification and Comparative Experiment of Micro Dynamic Colorimetric Method(mKC) for Detecting Bacterial Endotoxin Content in Antibody Drugs

目的:采用微量动态显色法(mKC)对抗体药物中的细菌内毒素含量进行检测,并对方法进行验证,同时与凝胶法的检测结果进行对比。方法:使用两个厂家的微量动态显色鲎试剂,对3个批号的样品,分别从标准曲线线性及范围、定量下限、检测限、干扰实验、精密度等方面进行方法学验证,同时进行微量动态显色法和凝胶法结果比对。结果:标准曲线相关系数的绝对值(|r|)均大于0.980,线性范围为10~0.1 EU/m L,定量下限为0.1 EU/m L,检测限为0.025 EU/m L,确定供试液的无干扰质量浓度为0.2 mg/m L,同一厂家试剂RSD小于25%,两个厂家试剂共计6次结果 RSD小于30%,微量动态显色法和凝胶法结果一致。结论:微量动态显色法可以用于抗体药物的细菌内毒素含量检测,并且可以替代传统的凝胶法用于抗体药物放行检测和稳定性考察。

Objective:To detect the content of bacterial endotoxin in antibody drugs by micro dynamic chromogenic assay(m KC),verify the method,and compare the results with gel method.Methods:The micro dynamic chromogenic limulus lysate from two manufacturers was used to verify the methodology of three batches of samples from the linearity and range of the standard curve,the lower limit of quantification,the detection limit,the interference experiment,and the precision.At the same time,the results of the micro dynamic chromogenic method and the gel method were compared.Results:The absolute value(|r|) of the correlation coefficient of the standard curve is greater than 0.980,the linear range is 10~0.1 EU/m L,the lower limit of quantification is 0.1 EU/m L,and the detection limit is0.025 EU/m L.It is determined that the non-interference mass concentration of the test solution is 0.2 mg/m L,and the RSD of the reagent from the same manufacturer is less than 25%.The RSD of six results of the reagents from two manufacturers is less than 30%.The results of the micro dynamic color method and the gel method are consistent.Conclusion:The micro dynamic color method can be used to detect the bacterial endotoxin content of antibody drugs,and can replace the traditional gel method for the release detection and stability inspection of antibody drugs.