TY - JOUR EP - 19896 SN - 2040-3364 TI - Multivalent nanobody engineering for enhanced physisorption and functional display on gold nanoparticles AV - public KW - Science & Technology KW - Physical Sciences KW - Technology KW - Chemistry KW - Multidisciplinary KW - Nanoscience & Nanotechnology KW - Materials Science KW - Multidisciplinary KW - Physics KW - Applied KW - Chemistry KW - Science & Technology - Other Topics KW - Materials Science KW - Physics KW - COLLOIDAL STABILITY KW - GENERATION KW - ANTIBODIES N1 - This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. N2 - The ease of expression and engineering of single domain antibodies, known as nanobodies, make them attractive alternatives to conventional antibodies in point-of-care diagnostics such as lateral flow assays. In lateral flow assays, gold nanoparticle bioconjugates serve as labels which display affinity molecules on the gold surface. While examples of nanobody gold nanoparticle bioconjugates exist, few utilise the simple one-step approach of physisorption owing to undesirable nanoparticle aggregation and loss of functionality. Here we show that engineering nanobodies into multivalent structures can significantly enhance their functionality when physisorbed onto gold nanoparticles. This approach enables resulting bioconjugates to withstand multiple processing steps required for long-term nanoparticle storage within lateral flow assays. Specifically, we show that the trivalent version of VHHV nanobody (VHH3) against the S1 protein of SARS-CoV-2 can be immobilised onto gold nanoparticles through passive adsorption. Unlike its monovalent and bivalent nanobody counterparts, using VHHV3 preserves nanoparticle stability under salt stress, blocking, washing, and freeze-drying conditions while maintaining picomolar sensitivity to the S1 protein. We anticipate that this facile strategy is a significant advancement towards the integration of nanobodies in lateral flow assay development. ID - discovery10199431 UR - http://dx.doi.org/10.1039/d4nr02762k Y1 - 2024/01/01/ JF - Nanoscale PB - ROYAL SOC CHEMISTRY A1 - Ayrton, John-Paul A1 - Ho, Chapman A1 - Zhang, Haoran A1 - Chudasama, Vijay A1 - Frank, Stefanie A1 - Thomas, Michael R VL - 16 SP - 19881 ER -