Nanobody engineering for SARS-CoV-2 neutralization and detection

• DOI: 10.1128/spectrum.04199-22
• Publication Date: 2024-02-16T14:00:21Z
• Authors: Liina Hannula; Suvi Kuivanen; Jonathan Lasham; Ravi Kant; Lauri Kareinen; Mariia Bogacheva; Tomas Strandin; Tarja Sironen; Jussi Hepojoki; Vivek Sharma; Petri Saviranta; Anja Kipar; Olli Vapalahti; Juha T. Huiskonen; Ilona Rissanen
• Keywords: Protein Engineering; Single-domain antibody

ABSTRACT In response to the ongoing COVID-19 pandemic, quest for coronavirus inhibitors has inspired research on a variety of small proteins beyond conventional antibodies, including robust single-domain antibody fragments, i.e., “nanobodies.” Here, we explore potential nanobody engineering in development antivirals and diagnostic tools. Through fusion domains that target distinct binding sites, engineered multimodular constructs neutralize wild-type SARS-CoV-2 Alpha Delta variants at high potency, with IC 50 values as low pM. Despite simultaneous epitopes, Beta Omicron were more resistant neutralization by nanobodies, which highlights importance accounting antigenic drift design biologics. To further applications outbreak management, present an assay based fusions nanobodies fragments NanoLuc luciferase can detect sub-nanomolar quantities spike protein single step. Our work showcases combat emerging infectious diseases. IMPORTANCE Nanobodies, binders derived from camelid antibody, are highly potent respiratory viruses offer several advantages over antibodies candidates specific therapies, stability production costs. this work, leverage unique properties apply them building blocks new therapeutic We report ultra-potent inhibition comprising multiple modules structure-guided combinations develop carry signal molecules, allowing rapid detection protein. results highlight effective countermeasures, both diagnostic, manage outbreaks viruses.