目的 对抗表皮生长因子2单克隆抗体的质量控制项目进行趋势分析。方法 利用BT-474细胞株作为靶细胞,通过细胞增殖抑制作用,使用CellTiter 96^AQ_ueous检测试剂盒进行抗表皮生长因子受体2单抗的生物学活性检测,以抗表皮生长因子受体2单抗参比品通过平行线分析的方法计算供试品效价;采用阳离子交换色谱方法,按面积归一化法计算主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值检测;通过对中国食品药品检定研究院(NIFDC)和企业质控实验室多批次抗表皮生长因子2单抗的检测结果,分别建立警戒限(±2s)和行动限(±3s),绘制趋势分析图,并对检测结果进行连续性及周期性趋势分析。结果 对2012~2014年某企业74批抗表皮生长因子2单抗的生物学活性检测结果显示,抗表皮生长因子受体2单抗供试品和参比品在生物学活性中均存在量效关系,在半对数坐标纸上呈平行的直线分布。生物学活性效价均值中国食品药品检定研究院与企业分别为(1.04±0.11)×10⁴和(0.94±0.08)×10⁴ U·mg^-1;蛋白质含量均值分别为(442.15±13.59)和(442.07±6.60) mg·瓶^-1;离子交换色谱主峰面积均值分别为(76.29±1.36)%和(73.76±1.17)%;pH值均值分别为(6.16±0.11)和(6.22±0.08)。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 首次通过对抗表皮生长因子受体2单克隆抗体的部分关键质量属性(CQA)趋势分析,反映了制品的变化情况,对评价单抗的批间一致性和生产工艺稳定性提供了参考,为其他单抗质量控制中关键质量属性的趋势分析提供重要指导和借鉴。

OBJECTIVE To analyze the trend in quality control of anti-HER2 monoclonal antibody.METHODS The biological activity of anti-HER2 monoclonal antibody (mAb) was monitored by the proliferation inhibition with CellTiter 96^AQ_ueousAssay kit using BT-474 cells as target, and the potency of anti-HER2 mAbs was calculated by comparison with the reference standard using the method of parallel analysis.Cation exchange chromatography method was used to detect the purity by calculating the area percentage of the main peak according to the area normalization method.Ultraviolet spectrophotometry was used to detect the protein content. And pH was detected by potential method. The warning limit(±2s)and action limit(±3s)were defined according to the determination results of several batches of anti-HERmAb by National Institutes for Food and Drug Control(NIFDC)and the quality control laboratory of manufacturer, based on which the trend graph was plotted, and the consistent and periodic trend of anti-HER2 mAb were analyzed.RESULTS The potencies of 74 batches of anti-HER2 antibodies from a certain enterprise during 2012 to 2014 were analyzed. The products and reference standard both showed a dose-response effect in biological activity, presenting parallel straight lines on semi-log coordinate paper.The potencies determined by NIFDC and the manufacturer were (1.04±0.11)×10⁴ and (0.94±0.08 )×10⁴ U·mg^-1, respectively. And the protein contents were (442.15±13.59) and(442.07± 6.60) mg·vial^-1.The main peak areas of IEC-HPLC were(76.29±1.36)% and (73.76±1.17)%, while the pHs were (6.16±0.11) and (6.22±0.08), 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 trend analysis of critical quality attributes (CQA) of anti-HER2 monoclonal antibody. These results reveal the quality changes of the products,which would help a lot in evaluating the batch consistency and production stability, and also provide reference for trend analysis of critical quality attributes for other monoclonal antibodies.