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重组人源单克隆抗体临床免疫原性检测方法的建立

发布时间:2018-03-10 22:55

  本文选题:重组人源单克隆抗体 切入点:rhuIgG- 出处:《中国生物制品学杂志》2016年07期  论文类型:期刊论文


【摘要】: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。
[Abstract]:Objective to establish a method for the detection of immunogenicity of recombinant human monoclonal antibody (rhu Ig G-04) in clinical study. Immunogenicity of monoclonal antibody rhu Ig G-04 in clinical studies, The method was optimized to remove the interference of rheumatoid factor (RF) by using fusion protein containing human Ig Gfc, and a large number of blank serum signal values and signal inhibition rates were calculated. The critical value of screening method and confirmatory method was determined. In the screening method, the normalized factor was determined to normalize the critical value of different batches. The tolerance of the detection method to rhu Ig G-04 in serum was analyzed. The optimal dilution ratio was 1 / 10, the optimal dilution was PBSs, the optimal dilution of second antibody Ig G-04-HRP was 1/10 000, the fusion protein with 1 mg/ml FC fragment in the second antibody could effectively remove the interference of RF. The normalization factor is 0.006, the critical value of batch detection is negative reference substance A normalized factor, and the concentration of drug in serum reaches 1 200 ng/ml, which has no effect on the results of screening method. The suitable concentration of rhu Ig G-04 was 100 渭 g / ml, and the critical value of confirmation method was 49 渭 g / ml. Conclusion the established immunogenicity detection method has high detection sensitivity and strong ability of anti-drug interference. The method of immunogenicity detection of human monoclonal antibody rhu Ig G-04 has been met with the basic requirements of antibody immunogenicity detection method, which provides a suitable method for the detection of immunogenicity of human monoclonal antibody rhu Ig G-04.
【作者单位】: 华北制药集团新药研究开发有限责任公司;
【基金】:“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)
【分类号】:R392-33

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