| 郭莎,张峰,刘春雨,于传飞,李萌,王文波,付志浩,俞小娟,王兰.一种抗TNFα单抗不同粒径级别蛋白聚体监测结果的比较与评价[J].中国药事,2018,32(8):1073-1078 |
| 一种抗TNFα单抗不同粒径级别蛋白聚体监测结果的比较与评价 |
| Comparison and Evaluation of Monitoring Results of Protein Aggregates with Different Orders of Magnitude of an Anti-TNFα Monoclonal Antibody |
| 投稿时间：2018-04-16 |
| DOI：10.16153/j.1002-7777.2018.08.011 |
| 中文关键词: 单克隆抗体 稳定性研究 蛋白聚体 分子排阻高效液相色谱法 微流数字成像技术 |
| 英文关键词: monoclonal antibody stability study protein aggregate size exclusion-high performance liquid chromatography microflow digital imaging |
| 基金项目:国家科技重大专项重大新药创制自身免疫疾病治疗的单抗类生物类似药质量评价和国家标准品建立（编号2018ZX09736008-006） |
| 作者 | 单位 | E-mail | | 郭莎 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 张峰 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 刘春雨 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 于传飞 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 李萌 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 王文波 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 付志浩 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 俞小娟 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | | | 王兰 | 中国食品药品检定研究院单克隆抗体产品室, 卫生部生物技术产品检定及标准化重点实验室, 北京 100050 | wanglan@nifdc.org.cn |
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| 中文摘要: |
| 目的：探讨在单克隆抗体（单抗）稳定性研究中，分子排阻高效液相色谱法（size exclusion-highperformance liquid chromatography，SEC-HPLC）、微流数字成像（microflow digital imaging，MDI）技术、澄清度及可见异物检查在不同粒径级别蛋白聚体监测中的交互作用。方法：分别采用SEC-HPLC、MDI、澄清度及可见异物检查研究肿瘤坏死因子α（TNFα）靶点单抗在4℃储存6个月、37℃和6000 lx光照条件下储存28 d后，蛋白聚体、不溶性微粒、澄清度及可见异物的不同变化。结果：SEC-HPLC结果表明，与样品在4℃储存6个月的结果相比，抗TNFα单抗的蛋白聚体含量在37℃和6000 lx照度处理28d后均增加均不明显。使用MDI技术可见，抗TNFα单抗经37℃和6000 lx照度处理28 d后，1~10 μm、10 μm以上、25 μm及以上粒径的微粒数量比在4℃储存6个月的样品有明显增多，且增多的微粒主要为蛋白聚体。通过目视法对样品的澄清度和可见异物检查发现，37℃储存和6000 lx照度处理28 d后的抗TNFα单抗澄清度低于4℃储存的样品；各储存条件下的抗TNFα单抗均未检出可见异物。结论：在单抗稳定性研究中，纳米级、微米级蛋白聚体的形成上并不具有一致性，因此，应采用多种方法对蛋白聚集情况进行表征和分析，避免单一方法的局限。 |
| 英文摘要: |
| Objective:To discuss the interaction of following methods in monitoring protein aggregate with different orders of magnitude, such as size exclusion-high performance liquid chromatography (SEC-HPLC), microflow digital imaging (MDI), clarity and visible foreign matter visual examination in the study of the stability of monoclonal antibodies (mAbs). Methods:SEC-HPLC MDI and visual examination were used separately to study the difference of protein aggregates, insoluble particles, clarity and visible foreign matter of an anti-tumor necrosis factor α (TNFα) mAb after being stored at 4℃ for 6 months or at 37℃ or under 6000 lx illumination for 28 days. Results:The results of SEC-HPLC showed that the proportion of protein aggregates of the anti-TNFα monoclonal antibody was not significantly increased after 28 days treatment at 37℃ and under 6000 lx illumination compared with the results of being stored at 4℃ for 6 months. The results of MDI showed that in the anti-TNFα monoclonal antibody after 28 days treatment at 37℃ or under 6000 lx illumination, the numbers in the particles with particle size of 1 to 10 μm, 10 μm and more than 25 μm were significantly higher than those in the samples stored at 4℃ for 6 months. And the increased particles are mainly protein aggregates. The clarity and visible foreign matter inspection of the sample by visual examination revealed that the clarity of the anti-TNFα monoclonal antibody after 28 days treatment at 37℃ or under 6000 lx illumination was less than that in the samples stored at 4℃. No visible particles were found in anti-TNFα samples under three different storage conditions. Conclusion:There was no consistency in the formation of nano-grade and micron-grade protein aggregates in monoclonal antibody stability study. Therefore, various methods should be used to characterize and analyze the protein aggregation in order to avoid limitation of a single method. |
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