Chaudhuri Samata, Korten Till, Diez Stefan
B CUBE - Center for Molecular Bioengineering, Technische Universität Dresden , 01069 Dresden, Germany.
Max Planck Institute of Molecular Cell Biology and Genetics , 01307 Dresden, Germany.
Bioconjug Chem. 2017 Apr 19;28(4):918-922. doi: 10.1021/acs.bioconjchem.7b00118. Epub 2017 Mar 14.
Engineering cargo-loading strategies is crucial to developing nanotechnological applications of microtubule-based biomolecular transport systems. Here, we report a highly efficient and robust bioconjugation scheme to load antibodies to microtubules. Our method takes advantage of the inverse-electron-demand Diels-Alder addition reaction between tetrazine and trans-cyclooctene: the fastest known bioorthogonal reaction, characterized by its excellent selectivity and biocompatibility. As proof of concept, we performed kinesin-1 gliding motility assays with antibody-conjugated microtubules and demonstrated the highly sensitive detection of fluorescent protein analyte down to 0.1 pM in microliter sample volumes. Importantly, the detection selectivity was retained in the presence of other fluorescent background proteins. We envision the applicability of our fast, simple, and robust conjugation method to a wide range of biosensing platforms based on biomolecular transport systems.
设计货物装载策略对于开发基于微管的生物分子运输系统的纳米技术应用至关重要。在此，我们报告了一种高效且稳健的生物共轭方案，用于将抗体装载到微管上。我们的方法利用了四嗪与反式环辛烯之间的逆电子需求狄尔斯-阿尔德加成反应：这是已知最快的生物正交反应，具有出色的选择性和生物相容性。作为概念验证，我们用抗体偶联的微管进行了驱动蛋白-1滑行运动测定，并证明在微升样品体积中能够高度灵敏地检测低至0.1 pM的荧光蛋白分析物。重要的是，在存在其他荧光背景蛋白的情况下，检测选择性得以保留。我们设想我们这种快速、简单且稳健的共轭方法可应用于基于生物分子运输系统的广泛生物传感平台。
