Robert Frederick Smith School of Chemical and Biomolecular Engineering , Cornell University , 113 Ho Plaza , Ithaca , New York 14850 , United States.
Department of Chemical and Biomolecular Engineering , University of Illinois at Urbana-Champaign , 600 S Mathews Ave , Urbana , Illinois 61801 , United States.
Bioconjug Chem. 2019 Sep 18;30(9):2452-2457. doi: 10.1021/acs.bioconjchem.9b00522. Epub 2019 Aug 21.
Site-specific modification of native antibodies has proven advantageous, as it enhances the properties of antibody-based bioconjugates without the need to manipulate the genetic code. However, native antibody modification is typically limited to strategies that introduce a single functional handle. In this work, we addressed this limitation by designing heterobifunctional substrates for microbial transglutaminase (MTG) that contain both azide and methyltetrazine "click" handles. Structure-conjugation relationships for these substrates were evaluated using the Her2-targeted antibody trastuzumab. Förster resonance energy transfer (FRET) was used to demonstrate that these chemical handles are mutually orthogonal. This orthogonality was leveraged for the one-pot synthesis of a bifunctional antibody-drug conjugate (ADC). This ADC, containing a maytansine-derived payload and a hydrophobicity-masking polyethylene glycol (PEG) side chain, demonstrated potent in vitro activity in SKOV3 cells. These studies establish the dual "click" approach as a powerful technique in the toolbox for native antibody modification.
通过对天然抗体进行定点修饰，可以在不改变遗传密码的情况下增强抗体结合物的性能，事实证明这是非常有利的。然而，天然抗体的修饰通常仅限于引入单个功能接头的策略。在这项工作中，我们通过设计含有叠氮化物和甲基四嗪“点击”接头的微生物转谷氨酰胺酶（MTG）的杂双功能底物来解决这一限制。使用曲妥珠单抗（靶向 Her2 的抗体）评估了这些底物的结构-连接关系。利用Förster 共振能量转移（FRET）证明了这些化学接头是相互正交的。这种正交性被用于一锅法合成双功能抗体药物偶联物（ADC）。该 ADC 含有美坦辛衍生的有效载荷和疏水性掩蔽的聚乙二醇（PEG）侧链，在 SKOV3 细胞中表现出很强的体外活性。这些研究确立了双“点击”方法是天然抗体修饰工具箱中的一种强大技术。
