摘要
目的对一种用于治疗人类表皮生长因子受体2(HER2)阳性乳腺癌的抗人类表皮生长因子受体2人源化单克隆抗体靶向生物制剂的关键质量属性进行多维度评价研究。方法采用高效液相色谱、毛细管电泳、动态光散射、圆二色谱、差示扫描量热技术、液质联用技术、体外活性实验以及表面等离子共振(SPR)技术分别对抗人类表皮生长因子受体2抗体的纯度、杂质、结构、活性等关键质量属性进行多维度分析。结果该抗体的单体纯度>98%,聚体杂质<2%;免疫球蛋白轻重链电泳纯度>97%,非糖重链杂质<1%;与参比生物制剂可比的圆二扫描图谱、二级结构预测结果和去折叠温度,N-糖修饰位点、种类和各糖分布比例与参比生物制剂高度相似;BT474增殖抑制活性、抗体依赖细胞介导的细胞毒性作用活性、抗原抗体亲和力、Fc受体亲和力等活性相关质量属性与参比生物制剂均无显著差异。结论该抗体高纯度、低杂质特性暗示其产品均一性和较低的安全风险,与参比生物制剂高度相似的高级结构、糖基化修饰和生物功能也从多个角度得到证实,这些正向质量评价是开展临床实验研究的前提,同时从某种程度上可以与临床安全和疗效指针建立关联。
OBJECTIVE To perform a multidimensional study on the critical quality attributes of a therapeutic anti-HER2 humanized monoclonal antibody targeting HER2-positive breast cancer. METHODS The critical quality attributes such as purity,impurities,structure,and function were evaluated using multidimensional analytical techniques including HPLC,CE-SDS,dynamic light scattering,circular dichroism,differential scanning calorimeter,LC-MS / MS,in vitro bioactivity assay and SPR binding kinetics. RESULTS The monomer content of the antibody was more than 98% while the polymer impurities were less than 2%. The sum of heavy chain and light chain peak area was over 97% while the non-glycosylated heavy chain impurities area was less than 1% on the CE-SDS electrophoretogram. The antibody drug demonstrated comparable CD spectrum,predicted secondary structure and unfolding temperatures to the reference product. It had highly similar N-glycan profile involving glycosylation site and N-glycan types to the reference product. The analysis showed no significant difference in the functional CQAs like BT474 proliferation inhibiting bioactivity,ADCC efficacy,the binding affinity to HER2 and Fc receptors between the evaluated drug product and the reference. CONCLUSION The characteristics of the monoclonal antibody drug such as high purity and few impurities indicate homogeneity and low-risk of safety issues. High similarity to the reference has been verified in multiple aspects like higher structure,glycosylation and function. The positive quality evaluation result is a prerequisite for clinical research. Moreover,to some extent,there can be potential correlation between CQAs and clinical safety and efficacy.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2015年第12期1054-1061,共8页
Chinese Pharmaceutical Journal
作者简介
王晓闻，女，硕士研究方向：抗体药物质量评价分析
通讯作者：谭青乔，男，博士 研究方向：抗体药物质量评价分析 Tel：（021）60757923 E-mail：tangqingqiao@sohu．com
