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Binding of monoclonal antibodies isolated from SARS-CoV-2 breakthrough infections to bat sarbecovirus RBDs. Nazia Thakur (9904162) Joseph Newman (5638340) Dongchun Ni (15188356) Jeffrey Seow (9415776) Abigail L. Hay (25157576) Yeonjae Lee (12256119) Ayush Upadhyay (25157573) Babatunde E. Ekundayo (25157578) John A. Hammond (7431785) Thomas P. Peacock (6536120) Kelvin Lau (1847764) Katie J. Doores (346210) Dalan Bailey (193037) Microbiology Cell Biology Evolutionary Biology Environmental Sciences not elsewhere classified Ecology Immunology Biological Sciences not elsewhere classified Cancer Infectious Diseases Virology multiple bat coronaviruses g ., rc g ., civet clade 1 viruses restricted ace2 usage mammalian ace2 proteins g ., rhgb07 using monoclonal antibodies clade 1 sarbecoviruses div >< p determined host range converting enzyme 2 2 convalescent individuals ace2 ), via using binding assays antigenicity using sera related bat sarbecoviruses using sarbecoviruses host range clade 3 pangolin ), neutralising antibodies entry assays binding domain sarbecoviruses interact sarbecoviruses exhibited rodent ace2 like rhgb07 generalist ace2 related antigenicity zoonotic potential vitally important still evidence specialist </ shifting pattern robust evidence research indicates racoon dog novel structure largely maintained including human immunodominant target human primate generalist phenotypes generalist </ especially omicron entry phenotypes diverse panel continued evolution breakthrough infections (A) Neutralisation of SARS-CoV-2 and Rs4231 RBD chimeras with SARS-CoV-2 RBD-specific monoclonal antibodies (mAb) (IC50 µg/ml), with dotted lines linking matched samples. The p values are shown, calculated by comparing the change in mean IC50 value and by using Tukey’s multiple comparison tests, respectively. (B) Spike structure of SARS-CoV-2 with 2 RBD-down and 1 RBD-up (PDB 7QTI) with the binding epitope regions of the different mAb competition groups is shown with both a side- and top-view (Group 1 = purple, Group 2 = orange, Group 3 = red, Group 3.5 = blue, Group 4 = green, Group 8 = non-RBD, NTD-specific, defined previously [<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003944#pbio.3003944.ref046" target="_blank">46</a>–<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003944#pbio.3003944.ref048" target="_blank">48</a>]. (C) SARS-CoV-2 RBD structures (PDB 7QTI) highlighting a broad epitope footprint of the mAb competition groups corresponding binding data. Each of the plots shows a single mAb binding in an ELISA using purified (C) SARS-CoV-2 or (E) other bat sarbecovirus RBDs. Plots show raw optical density (OD) values with curves calculated using a least-squares regression non-linear fit. (D) Epitope binning by surface plasmon resonance (SPR) identifies 5 epitope communities broadly aligning with the antibody competition groups. Grey circles indicate communities by setting the associated dendrogram threshold to 0.45. Solid lines indicate symmetrical blocking, regardless of which mAb is the ligand. Dotted lines indicate asymmetrical blocking, with the arrowhead indicating the direction of blocking, pointing towards the antibody immobilised as a ligand. Circles indicate antibodies for which ligand and analyte data was available, squares indicate only analyte data was available. The data underlying this Figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003944#pbio.3003944.s017" target="_blank">S1 Data</a>. 2026-10-01T17:40:07Z Image Figure 10.1371/journal.pbio.3003944.g006 https://figshare.com/articles/figure/_p_Binding_of_monoclonal_antibodies_isolated_from_SARS-CoV-2_breakthrough_infections_to_bat_sarbecovirus_RBDs_p_/34047382 CC BY 4.0