Department of Virology and Molecular Biology, Wageningen Bioveterinary Research, Lelystad, The Netherlands.
Laboratory of Virology, Wageningen University, Lelystad, The Netherlands.
Methods Mol Biol. 2022;2446:313-326. doi: 10.1007/978-1-0716-2075-5_15.
Multimerization of single-domain antibodies (sdAbs) is instrumental for construction of antibody molecules with high avidity, extended in vivo half-life, and tailor-made biological activity. Two-component superglues, based on bacterium-derived peptides (Tags) and small protein domains (Catchers) that form isopeptide bonds when in close proximity, enable the creation of multimers by simply mixing of the individual components. Here, we provide detailed methods for the construction of sdAbs and scaffolds bearing genetically fused superglue components and their assembly into multimeric complexes.
多聚化单域抗体（sdAb）对于构建具有高亲和力、延长体内半衰期和定制生物活性的抗体分子至关重要。由细菌衍生肽（Tags）和小蛋白结构域（Catchers）组成的双组分超级胶水在近距离时形成异肽键，通过简单混合单个成分即可实现多聚体的构建。在此，我们提供了构建携带遗传融合超级胶水组件的 sdAb 和支架的详细方法，以及将它们组装成多聚体复合物的方法。
