英夫利西单抗质量控制中的趋势分析

目的：对英夫利西单抗的质量控制项目进行趋势分析。方法：利用WEHI164-13var细胞株作为靶细胞，通过细胞增殖抑制作用，使用CellTiter 96AQ_ueous检测试剂进行英夫利西单抗的生物学活性检测，以英夫利西单抗参比品计算供试品相对百分效价；采用分子排阻色谱方法，按面积归一化法计算单体峰面积百分比进行抗体纯度检测；采用紫外分光光度法进行蛋白质含量检测；采用电位法进行pH检测；对中国食品药品检定研究院（NIFDC）和企业质控实验室多批次英夫利西单抗的上述检测结果，分别建立警戒限（均值±2SD）和行动限（均值±3SD），绘制趋势分析图，对结果进行连续性趋势分析；并对生物学活性的检测结果进行周期性趋势分析。结果：英夫利西单抗供试品和参比品在生物学活性中均存在量效关系，符合四参数方程式：y=（A-D）/［1+（x/C）^B］+D，在半对数坐标纸上呈S型曲线分布。对2008—2017年某企业65批英夫利西单抗的生物学活性检测结果显示，生物学活性相对百分效价均值NIFDC与企业分别为（98.000±9.384）%和（103.046±4.421）%；蛋白质含量均值分别为（96.431±3.000）mg·瓶^-1和（99.692±1.185）mg·瓶^-1；分子排阻色谱单体峰面积均值分别为（99.771±0.177）%和（99.543±0.179）%；pH均值分别为7.272±0.111和7.309±0.038。对上述结果进行连续性和周期性趋势分析，总体趋势较平稳。结论：通过对英夫利西单抗的部分关键质量属性（CQA）进行趋势分析，结果反映了该制品的变化情况，对评价英夫利西单抗的批间一致性和生产工艺稳定性提供了参考，也为其他单抗质控中的趋势分析提供重要指导。

Objective:To carry out trend analysis in the project of quality control of infliximab. Methods:The WEHI164-13 var cell line was used as the target cell,the inhibition of cell proliferation was used to detect the biological activity of infliximab using the CellTiter 96AQ_ueous detection reagent,and the relative potency was calculated by EC₅₀ (reference)/EC₅₀ (test article)×100%. Size exclusion chromatography methods was used to detect the purity by calculating the area percentage of the monomer peak according to the area normalization method. Ultraviolet spectrophotometry was used to detect the protein content. Potential method was used to detect pH. The warning limits (mean±2SD)and action limit (mean±3SD)were established according to the test results of several batches of infliximab by National Institutes for Food and Drug Control (NIFDC) and the quality control laboratory of manufacturer. Trend analysis graph was plotted and the consistency trend of infliximab was analyzed. Finally,the results of biological activity were carried out periodic trend analysis. Results:Both the samples and reference showed a dose-response relationship in biological activity,in line with the four-parameter equation:y=(A-D)/[1+(x/C)^B]+D,presenting S-shaped curve distribution on semi-log coordinate paper. The results of the biological activity test of 65 batches of infliximab in a certain enterprise from 2008 to 2017 showed that the average relative potency of biological activity of the NIFDC and the enterprise were (98.000±9.384)% and (103.046±4.421)%,respectively. And mean values of the protein content were (96.431±3.000)mg per vial and (99.692±1.185)mg per vial respectively. The monomer peak area of SEC-HPLC were (99.771±0.177)% and (99.543±0.179)%,while pH were 7.272±0.111 and 7.309±0.038 respectively. The consistent and periodic trend analysis of the above-mentioned results showed that the total trend was relatively steady. Conclusion:This is the first time to conduct the trend analysis in some critical quality attributes (CQA) of infliximab. These results reveal the changes of the products,which provide references for evaluating the batch consistency and production stability,and also provide important guides for trend analysis in quality control of other monoclonal antibodies. 

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