eprintid: 10199431 rev_number: 9 eprint_status: archive userid: 699 dir: disk0/10/19/94/31 datestamp: 2024-11-04 11:38:09 lastmod: 2024-11-04 11:38:09 status_changed: 2024-11-04 11:38:09 type: article metadata_visibility: show sword_depositor: 699 creators_name: Ayrton, John-Paul creators_name: Ho, Chapman creators_name: Zhang, Haoran creators_name: Chudasama, Vijay creators_name: Frank, Stefanie creators_name: Thomas, Michael R title: Multivalent nanobody engineering for enhanced physisorption and functional display on gold nanoparticles ispublished: pub divisions: UCL divisions: B04 divisions: C06 divisions: F47 divisions: F56 divisions: F64 keywords: Science & Technology, Physical Sciences, Technology, Chemistry, Multidisciplinary, Nanoscience & Nanotechnology, Materials Science, Multidisciplinary, Physics, Applied, Chemistry, Science & Technology - Other Topics, Materials Science, Physics, COLLOIDAL STABILITY, GENERATION, ANTIBODIES note: This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. abstract: 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. date: 2024-01-01 date_type: published publisher: ROYAL SOC CHEMISTRY official_url: http://dx.doi.org/10.1039/d4nr02762k oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 2327559 doi: 10.1039/d4nr02762k medium: Print-Electronic lyricists_name: Thomas, Michael lyricists_name: Chudasama, Vijay lyricists_name: Frank, Stefanie lyricists_name: Ho, Chapman lyricists_id: MTHOA12 lyricists_id: VCHUD30 lyricists_id: SFRAN44 lyricists_id: CHOXX78 actors_name: Thomas, Michael actors_id: MTHOA12 actors_role: owner funding_acknowledgements: EP/T517793/1 [EPSRC]; EP/R00529X/1 [EPSRC IRC in Agile Early Warning Sensing Systems for Infectious Diseases and Antimicrobial Resistance grant]; EP/Y530542/1 [DHSC/EPSRC] full_text_status: public publication: Nanoscale volume: 16 pagerange: 19881-19896 pages: 16 event_location: England issn: 2040-3364 citation: Ayrton, John-Paul; Ho, Chapman; Zhang, Haoran; Chudasama, Vijay; Frank, Stefanie; Thomas, Michael R; (2024) Multivalent nanobody engineering for enhanced physisorption and functional display on gold nanoparticles. Nanoscale , 16 pp. 19881-19896. 10.1039/d4nr02762k <https://doi.org/10.1039/d4nr02762k>. Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10199431/7/Thomas_d4nr02762k.pdf