刘艳玲;曹晨华;赵伟;刘晓志;段迎霞;高健;

• 1:华北制药集团新药研究开发有限责任公司

摘要(Abstract)：

目的建立重组人源单克隆抗体rhu Ig G-04临床研究中免疫原性检测方法。方法建立由筛选法(桥连ELISA法)和确证法(免疫清除法)组成的多重检验方法检测重组人源单克隆抗体rhu Ig G-04临床研究中的免疫原性,并对方法进行优化,利用含有人Ig G(Fc)的融合蛋白去除类风湿因子(rheumatoid factor,RF)的干扰;通过统计大量空白血清信号值及信号抑制率,确定筛选法和确证法的临界值。在筛选法中确定了归一化因子对不同批次的临界值进行归一化处理。分析检测法对血清中rhu Ig G-04的耐受性。结果筛选法中,样品最佳稀释倍数为1/10,最佳稀释液为PBS,二抗rhu Ig G-04-HRP的最适稀释度为1/10 000;在二抗中加入浓度为1 mg/ml Fc片段的融合蛋白能有效去除RF的干扰;归一化因子为0.006,批检测临界值为阴性对照品A值+归一化因子;血清中药物浓度达到1 200 ng/ml时,对筛选法结果无影响。确证法中,向样品中加入rhu Ig G-04的适宜浓度为100μg/ml,确证法临界值为49%。结论建立的免疫原性检测方法具有较高的检测灵敏度,抗药物干扰能力强,达到了抗体药物免疫原性检测方法的基本要求,为人源单抗rhu Ig G-04免疫原性的检测提供了适宜的手段。

关键词(KeyWords)： 重组人源单克隆抗体;rhuIgG-04;免疫原性;酶联免疫吸附测定

Abstract:

Keywords:

基金项目(Foundation): “重大新药创制”国家科技重大专项创新型人源单抗药物研制平台及重大品种开发(2014ZX09201041)

作者(Authors): 刘艳玲;曹晨华;赵伟;刘晓志;段迎霞;高健;

DOI: 10.13200/j.cnki.cjb.001382

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