Nanobody in a Double “Y”-Shaped Assembly: A Promising Candidate for Lateral Flow Immunoassays

作者：Yumeng Xiao, Hongjie Dong, Cancan Wu, Kundi Zhang, Xiaoqiong Jiang, Junyu Chen, Hongwei Wang, Sujuan Xu, Fengyu Zhang, Lichuan Gu · 发表于：Analytical Chemistry · 年份：2024 · DOI：10.1021/acs.analchem.4c00509 · 被引用次数：15 · 研究领域：Monoclonal and Polyclonal Antibodies Research、Advanced Biosensing Techniques and Applications、Biosensors and Analytical Detection

Derived from camelid heavy-chain antibodies, nanobodies (Nbs) are the smallest natural antibodies and are an ideal tool in biological studies because of their simple structure, high yield, and low cost. Nbs possess significant potential for developing highly specific and user-friendly diagnostic assays. Despite offering considerable advantages in detection applications, knowledge is limited regarding the exclusive use of Nbs in lateral flow immunoassay (LFIA) detection. Herein, we present a novel double "Y" architecture, achieved by using the SpyTag/SpyCatcher and Im7/CL7 systems. The double "Y" assemblies exhibited a significantly higher affinity for their epitopes, as particularly evident in the reduced dissociation rate. An LFIA employing double "Y" assemblies was effectively used to detect the severe acute respiratory syndrome coronavirus-2 N protein, with a detection limit of at least 500 pg/mL. This study helps broaden the array of tools available for the development of Nb-based diagnostic techniques.