Craenmehr Fenna W B, Gräwe Alexander, Veenbrink Victor A, Bellan Riccardo, Merkx Maarten, Dankers Patricia Y W
Institute for Complex Molecular Systems, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.
Department of Biomedical Engineering, Laboratory of Chemical Biology, Eindhoven University of Technology, Eindhoven 5600 MB, The Netherlands.
Bioconjug Chem. 2025 Jun 18;36(6):1197-1207. doi: 10.1021/acs.bioconjchem.5c00046. Epub 2025 May 14.
Supramolecular assemblies hold great potential as biomaterials for several biomedical applications. The modification of supramolecular biomaterials is needed to achieve controlled bioactive functions. Supramolecular ureidopyrimidinone (UPy) monomers have been shown to assemble into long supramolecular polymers that can be functionalized with bioactive peptides and visualized as UPy-fibers. So far, the introduction of biological functionality has been limited to small molecules and peptides. Here, we describe a general method based on SpyTag-SpyCatcher chemistry for conjugating full-length proteins with biologically relevant functions to μm-long UPy fibers via native peptide bond formation, yielding 100% conversion in a 5:95 mol % coassembly of UPy-SpyTag with UPy-glycinamide. The conjugation of monoclonal antibodies is performed using photo-cross-linkable protein G domains. We demonstrate intact fibers and colocalization of antibodies and UPy-fibers using biophysical and imaging methods and achieve recruitment of supramolecular assemblies to the surface of mammalian cells via the EGFR-specific antibody Cetuximab. The approach introduced here represents a robust and widely applicable postassembly modification method that shows promise in the functionalization of future biomaterials.
超分子组装体作为用于多种生物医学应用的生物材料具有巨大潜力。为实现可控的生物活性功能，需要对超分子生物材料进行修饰。超分子脲基嘧啶酮（UPy）单体已被证明可组装成长的超分子聚合物，这些聚合物可用生物活性肽进行功能化，并可视化为UPy纤维。到目前为止，生物功能的引入仅限于小分子和肽。在此，我们描述了一种基于SpyTag-SpyCatcher化学的通用方法，用于通过天然肽键形成将具有生物学相关功能的全长蛋白质与微米长的UPy纤维偶联，在UPy-SpyTag与UPy-甘氨酰胺以5:95摩尔%共组装时实现100%的转化率。单克隆抗体的偶联使用可光交联的蛋白G结构域进行。我们使用生物物理和成像方法证明了纤维的完整性以及抗体与UPy纤维的共定位，并通过表皮生长因子受体（EGFR）特异性抗体西妥昔单抗实现了超分子组装体在哺乳动物细胞表面的募集。本文介绍的方法代表了一种强大且广泛适用的组装后修饰方法，在未来生物材料的功能化方面显示出前景。
