Department of Protein Chemistry , Genentech, Inc. , 1 DNA Way , South San Francisco , California 94080 , United States.
Department of Pathology , Genentech, Inc. , 1 DNA Way , South San Francisco , California 94080 , United States.
Bioconjug Chem. 2018 Jul 18;29(7):2468-2477. doi: 10.1021/acs.bioconjchem.8b00362. Epub 2018 Jun 13.
Despite the recent success of antibody-drug conjugates (ADCs) in cancer therapy, a detailed understanding of their entry, trafficking, and metabolism in cancer cells is limited. To gain further insight into the activation mechanism of ADCs, we incorporated fluorescence resonance energy transfer (FRET) reporter groups into the linker connecting the antibody to the drug and studied various aspects of intracellular ADC processing mechanisms. When comparing the trafficking of the antibody-FRET drug conjugates in various different model cells, we found that the cellular background plays an important role in how the antigen-mediated antibody is processed. Certain tumor cells showed limited cytosolic transport of the payload despite efficient linker cleavage. Our FRET assay provides a facile and robust assessment of intracellular ADC activation that may have significant implications for the future development of ADCs.
尽管抗体药物偶联物（ADC）在癌症治疗方面最近取得了成功，但对它们在癌细胞中的进入、转运和代谢的详细了解仍有限。为了更深入地了解 ADC 的激活机制，我们将荧光共振能量转移（FRET）报告基团整合到连接抗体和药物的连接子中，并研究了细胞内 ADC 处理机制的各个方面。在比较不同模型细胞中抗体-FRET 药物偶联物的转运时，我们发现细胞背景在抗原介导的抗体处理中起着重要作用。尽管有效切割连接子，但某些肿瘤细胞显示出有效切割连接子，但细胞溶质转运有限。我们的 FRET 测定法为细胞内 ADC 激活提供了一种简单而强大的评估方法，这可能对 ADC 的未来发展具有重要意义。
