Site-specific labeling of an anti-MUC1 antibody: probing the effects of conjugation and linker chemistry on the internalization process.
Site-specific labeling of an anti-MUC1 antibody: probing the effects of conjugation and linker chemistry on the internalization process.
复制标题
抗 MUC1 抗体的位点特异性标记：探讨缀合和连接化学对内化过程的影响
DOI：
10.1039/c8ra09902b
复制
发表时间：
2019-01-14
期刊：
影响因子：
3.9
通讯作者：
Jiang, Xianxing
中科院分区：
文献类型：
--
作者：
Xu, Hongjiao;Gan, Lu;Han, Ying;Da, Yifan;Xiong, Jiale;Hong, Sihua;Zhao, Qian;Song, Nazi;Cai, Xiaoqing;Jiang, Xianxing
抗体-药物缀合物（ADC）作为一种有吸引力的癌症治疗方法最近受到了极大的关注。尽管ADC设计被认为对于ADC的相对功效是重要的，但ADC的结构特征如何影响分子的内化过程和细胞内运输仍然未被探索。在此，我们报告我们的努力，在设计抗体为基础的代理商的共轭和接头化学的细胞内吞作用的影响。设计了一系列抗MUC 1单链可变片段（scFv-SM 3）缀合物，其具有独特的结构特征，包括缀合方法、连接位点和接头化学。体外共聚焦成像显示，随机赖氨酸缀合和位点特异性缀合（包括C-末端修饰或内部位点缀合）都可以提供对靶表达细胞具有相似结合亲和力和细胞摄取的抗体缀合物。时程内化研究表明，具有短聚乙二醇接头的SM 3-缀合物胜过那些缺乏任何亲水性接头的SM 3-缀合物，以获得更高的细胞摄取和更快的内化速率。还使用MUC 1过表达肿瘤细胞在小鼠异种移植模型中测试了具有最高亲和力和内化率的SM 3缀合物。我们的研究结果表明，接头和缀合化学在抗体缀合物的内化过程中起着重要作用，这反过来又会影响ADC的治疗效果。
Antibody-drug conjugates (ADCs) have recently received enormous attention as an attractive approach for cancer therapy. Although ADC design has been believed to be important for the relative efficacy of ADCs, it remains unexplored how the structural characteristics of ADCs would impact the internalization process and intracellular trafficking of the molecules. Herein, we report our efforts in investigating the cellular endocytosis implications of the conjugation and linker chemistry in designing antibody-based agents. A series of anti-MUC1 single-chain variable fragment (scFv-SM3) conjugates were designed with unique structural characteristics ranging from conjugation methods, sites of attachment and linker chemistry. In vitro confocal imaging showed that both random lysine-conjugation and site-specific conjugation, including C-terminus modification or internal site conjugation, could afford antibody conjugates with similar binding affinity and cellular uptake to target-expressing cells. Time-course internalization studies demonstrated that SM3-conjugates with short polyethylene glycol linkers outcompeted those that lack any hydrophilic linkers for higher cellular uptake and faster internalization rate. The SM3-conjugates with the highest affinity and internalization rate were also tested in mouse xenograft models using MUC1-overexpressing tumor cells. Our results indicate that the linker and conjugation chemistry play an important role in the internalization process of antibody conjugates, and this in turn could impact the therapeutic effects of ADCs.
登录
查看更多内容
影响因子：
5.5
作者：
Wals K;Ovaa H
通讯作者：
Ovaa H
抗MUC1特异性抗体的上升和下降。
影响因子：
3.7
作者：
Thie H;Toleikis L;Li J;von Wasielewski R;Bastert G;Schirrmann T;Esteves IT;Behrens CK;Fournes B;Fournier N;de Romeuf C;Hust M;Dübel S
通讯作者：
Dübel S
影响因子：
14.8
作者：
通讯作者：
--
DOI：
10.1002/anie.201509432
发表时间：
2016-02-12
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Fang T;Duarte JN;Ling J;Li Z;Guzman JS;Ploegh HL
通讯作者：
Ploegh HL
影响因子：
78.5
作者：
Kufe, Donald W.
通讯作者：
Kufe, Donald W.