biostudies-literatureDong HThe Hong Kong Research Grants Council Theme-Based Research SchemeNational Natural Science FoundationHong Kong Research Grants Council Collaborative Research FundThe Health@InnoHK, Innovation and Technology Commission of Hong KongThe Hong Kong Health and Medical Research FundThe Emergency Key Program of Guangzhou LaboratoryWanke Scientific Research ProgramNational Key Plan for Scientific Research and Development of Chinae1012625https://www.ebi.ac.uk/biostudies/studies/S-EPMC11554226biostudies-literatureUnknown20(11)The ongoing evolution and immune escape of SARS-CoV-2, alongside the potential threat of SARS-CoV-1 and other sarbecoviruses, underscore the urgent need for effective strategies against their infection and transmission. This study highlights the discovery of nanobodies from immunized alpacas, which demonstrate exceptionally broad and potent neutralizing capabilities against the recently emerged and more divergent SARS-CoV-2 Omicron subvariants including JD.1.1, JN.1, KP.3, KP.3.1.1, as well as SARS-CoV-1 and coronaviruses from bats and pangolins utilizing receptor ACE2. Among these, Tnb04-1 emerges as the most broad and potent, binding to a conserved hydrophobic pocket in the spike's receptor-binding domain, distinct from the ACE2 binding site. This interaction disrupts the formation of a PLoS pathogensSuper broad and protective nanobodies against Sarbecoviruses including SARS-CoV-1 and the divergent SARS-CoV-2 subvariant KP.3.1.1.PMC11554226C7156-20G321712022022YFC2303400, 2023YFC304330082241072T11-709/21-NEKPG22-012021YFC0864500, 2022YFF1203100202210800562021YFC2300104 and 2022YFF120310382150205, 322709832022YFC260410092169205COVID1903010 and COVID190123Liu NLiang YYunfei Zeng IWang HZhang LWei ZGuo QShi XZhang QDong HChen ZChen YNiu MWang XWei JShan SWu YChen JZhou RZeng YLi MYang ZYang YZhu KYang QfalseSuper broad and protective nanobodies against Sarbecoviruses including SARS-CoV-1 and the divergent SARS-CoV-2 subvariant KP.3.1.1.The ongoing evolution and immune escape of SARS-CoV-2, alongside the potential threat of SARS-CoV-1 and other sarbecoviruses, underscore the urgent need for effective strategies against their infection and transmission. This study highlights the discovery of nanobodies from immunized alpacas, which demonstrate exceptionally broad and potent neutralizing capabilities against the recently emerged and more divergent SARS-CoV-2 Omicron subvariants including JD.1.1, JN.1, KP.3, KP.3.1.1, as well as SARS-CoV-1 and coronaviruses from bats and pangolins utilizing receptor ACE2. Among these, Tnb04-1 emerges as the most broad and potent, binding to a conserved hydrophobic pocket in the spike's receptor-binding domain, distinct from the ACE2 binding site. This interaction disrupts the formation of a 2024-01-01T00:00:00Z2024 Nov2026-07-01T03:15:46.25Z2025-04-03T23:55:42.023ZS-EPMC115542263952759410.1371/journal.ppat.1012625