毕华;李永红;韩春梅;秦玺;史新昌;裴德宁;丁有学;饶春明;

• 1:中国食品药品检定研究院

摘要(Abstract)：

目的建立牛痘病毒粒细胞巨噬细胞集落刺激因子(granulocyte macrophage colony stimulating factor,GM-CSF)注射液中GM-CSF活性及表达量的梯度分析法。方法采用TF-1细胞增殖法,四参数方程拟合检测不同浓度梯度样品的GM-CSF生物学活性;采用ELISA法,二次方程拟合检测不同浓度梯度样品的GM-CSF表达量。结果牛痘病毒GM-CSF注射液及标准品的浓度与GM-CSF的生物学活性呈量效依赖关系,随着GM-CSF浓度的增加,其GM-CSF的生物学活性增强,数据经四参数方程拟合后呈S型曲线,相关系数分别为0.953和0.997,将剂量反应曲线中直线部分的浓度对数和相应的吸光度值作直线回归,两回归线的斜率b之间差异无统计学意义(P>0.05);用该方法检测1批牛痘病毒GM-CSF注射液感染上清中GM-CSF活性,平均值为(181±16)GCU,变异系数为8.84%。牛痘病毒GM-CSF注射液及标准品的浓度与GM-CSF表达量呈量效依赖关系,随着稀释倍数的增加,GM-CSF的表达量也降低,采用二次方程进行拟合,r2均﹥0.99;用该方法检测1批牛痘病毒GM-CSF注射液感染上清中GM-CSF表达量,均符合表达量与感染浓度成正比升高的规律和趋势,且均>15 ng/m L。结论建立的方法稳定、可靠、适用性强,能表明病毒感染量与GM-CSF表达活性及表达量间的量效关系,对同类产品的质量控制具有借鉴意义。

关键词(KeyWords)： 牛痘病毒;粒细胞巨噬细胞集落刺激因子;生物学活性;表达量;基因治疗;质量控制

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 毕华;李永红;韩春梅;秦玺;史新昌;裴德宁;丁有学;饶春明;

DOI: 10.13200/j.cnki.cjb.002289

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