报告基因法测定聚乙二醇化Ⅰ型干扰素生物学活性的研究

摘要

目的 考察《中国药典》(2015年版)中报告基因法测定聚乙二醇化Ⅰ型干扰素生物学活性的方法适用性.方法 根据ICH指导原则,验证报告基因法的专属性、线性、准确性(回收率)、精密度(重复性、中间精密度)等指标,并对报告基因法与病毒抑制法的测定结果进行比较.结果 使用报告基因法测定聚乙二醇Ⅰ型干扰素活性具有良好的专属性,剂量反应关系符合四参数方程,回收率为85％～115％,重复性为6.66％～7.70％,不同测定日期、不同实验员和不同细胞代次的中间精密度分别为10.64％～ 11.01％、13.04％～15.12％和7.61％～ 8.54％,报告基因法与病毒抑制法的测定结果差异无统计学意义(P＞0.05).结论 报告基因法可以用于聚乙二醇化Ⅰ型干扰素生物学活性测定.%Objective To investigate the applicability of reporter gene assay in Chinese Pharmacopoeia (2015 edition) for determination of the bioactivity of polyethylene glycol (PEG) type I interferon.Methods According to ICH guidelines,specificity,linearity,accuracy (recovery) and precision (repeatability and intermediate precisions) of reporter gene assay were validated.The determination results of reporter gene assay and cytopathic inhibition assay were also compared.Results Application of reporter gene assay for determination of the bioactivity of PEG type Ⅰ interferon had good specificity,the dose-response of which was fitted to 4-parameter logistic model.The recovery rate was 85％-115％,the repeatability of the assay was 6.66％-7.70％.The intermediate precision of different dates,laboratory technicians and cell passages was 10.64％-11.01％,13.04％-15.12％ and 7.61％-8.54％ respectively.The difference of the determination results between reporter gene assay and cytopathic inhibition assay was not statistically significant (P ＞ 0.05).Conclusion The reporter gene assay is applicable for bioactivity determination of PEG type Ⅰ interferon.