El Hebieshy Angela F, Wijfjes Zacharias, Le Gall Camille M, Middelburg Jim, de Roode Kim E, Fennemann Felix L, Sluijter Marjolein, van Hall Thorbald, Dijkstra Douwe J, Trouw Leendert A, van Dalen Floris J, Rodgers Furones Andrea, van der Schoot Johan M S, Derksen Ian, de Haard Hans, van der Woning Bas, Talavera Ormeño Cami M P, van Doodewaerd Bjorn R, Figdor Carl G, van der Heden van Noort Gerbrand J, Parren Paul W H I, Heskamp Sandra, Ovaa Huib, Verdoes Martijn, Scheeren Ferenc A
Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, the Netherlands.
Department of Dermatology, Leiden University Medical Center, Leiden, the Netherlands.
Nat Biomed Eng. 2025 Apr 9. doi: 10.1038/s41551-024-01342-z.
Antibody conjugates are the foundation of a wide range of diagnostic and therapeutic applications. Although many antibody-conjugation techniques are robust and efficient, obtaining homogeneous multimeric conjugation products remains challenging. Here we report a modular and versatile technique for the site-directed multivalent conjugation of antibodies via the small-protein ubiquitin. Specifically, multiple ubiquitin fusions with antibodies, antibody fragments, nanobodies, peptides or small molecules such as fluorescent dyes can be conjugated to antibodies and nanobodies within 30 min. The technique, which we named 'ubi-tagging', allowed us to efficiently generate a bispecific T-cell engager as well as nanobodies conjugated to dendritic-cell-targeted antigens that led to potent T-cell responses. Using both recombinant ubi-tagged proteins and synthetic ubiquitin derivatives allows for the iterative, site-directed and multivalent conjugation of antibodies and nanobodies to a plethora of molecular moieties.
抗体偶联物是广泛的诊断和治疗应用的基础。尽管许多抗体偶联技术强大且高效，但获得均一的多聚体偶联产物仍然具有挑战性。在此，我们报告了一种通过小蛋白泛素进行抗体的位点定向多价偶联的模块化通用技术。具体而言，抗体、抗体片段、纳米抗体、肽或诸如荧光染料等小分子与多个泛素的融合物可在30分钟内与抗体和纳米抗体偶联。我们将该技术命名为“泛素标记”，它使我们能够高效地生成双特异性T细胞衔接子以及与树突状细胞靶向抗原偶联的纳米抗体，从而引发强烈的T细胞反应。使用重组泛素标记蛋白和合成泛素衍生物能够实现抗体和纳米抗体与大量分子部分的迭代、位点定向和多价偶联。
