目的: 对国产抗人类表皮生长因子受体2（HER2）单抗部分关键质量属性进行质量研究,并建立质量控制方法。方法: 根据曲妥珠的产品特性和作用机制,对质量属性进行评估分级,以7批原研曲妥珠为对照,检测7批国产抗HER2单抗部分关键质量属性。利用离子交换色谱检测电荷异质性;通过BT474细胞增殖抑制法检测生物学活性;利用生物膜干涉技术检测抗原抗体亲和力、单抗与FcRn亲和力以及利用表面等离子共振技术检测单抗与FcγRⅢa亲和力;利用差示扫描量热检测热稳定性。结果: 7批原研曲妥珠的离子交换色谱主峰面积百分比、生物学活性EC50值的Mean±3SD分别为（73.87±2.21）%、（0.20±0.06）μg·mL-1,7批国产抗HER2单抗相应检测结果分别为（73.54±3.24）%,（0.20±0.05）μg·mL-1。原研曲妥珠抗原抗体亲和力、单抗与FcγRⅢa亲和力及单抗与FcRn亲和力的平衡常数值（KD）分别为1×10-10、1×10-6、1×10-7,7批国产抗HER2单抗相应检测结果分别为1×10-10、1×10-6、1×10-7。国产抗HER2单抗与原研曲妥珠热稳定性图谱一致。结论: 本研究建立了抗HER2单抗关键质量属性研究方法,为该类国产单抗质量控制提供参考依据。
Objective: To study and establish a quality control method for some critical quality attributes of the domestic anti human epidermal growth factor receptor 2 (HER2) monoclonal antibodies. Methods: Based on the product characteristics and effects mechanism of trastuzumab, the quality attributes were evaluated and graded. And some critical quality attributes of 7 batches of the domestic anti-HER2 monoclonal antibody were detected using 7 batches of reference trastuzumabs as the control. The charge heterogeneity was analyzed by ion exchange-high performance liquid chromatography (IEC-HPLC). Biological activity was determined by BT474 cell proliferation inhibition assay. Bio-Layer Interferometry (BLI) was used to measure antigen-antibody affinity, monoclonal antibody affinity with FcRn, and Surface Plasmon Resonance (SPR) was used to measure the affinity of monoclonal antibody with FcγRⅢa. Thermal stability was evaluated by Differential Scanning Calorimetry (DSC). Results: The mean±3SD values of IEC main peak areas percent and EC50 of 7 batches of original trastuzumabs were (73.87±2.21)% and (0.20±0.06) μg·mL-1, respectively. The results of the domestic anti HER2 monoclonal antibodies were (73.54±3.24)%, (0.20±0.05) μg·mL-1, respectively. The KD values of antigenantibody affinity, antibody-FcγRⅢa affinity and antibody-FcRn affinity were 1×10-10, 1×10-6, 1×10-7, respectively. The results of 7 batches of the domestic anti-HER2 monoclonal antibodies were 1×10-10, 1×10-6, 1×10-7, respectively. The thermal stability mappings of the domestic anti-HER2 monoclonal antibodies were consistent with those of the original trastuzumabs. Conclusion: A method for quality study of anti-HER2 monoclonal antibody is developed, which provides a reference for quality control of this class domestic monoclonal antibody.
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