tetano
Editor, Senior Moderator
Proc Natl Acad Sci U S A
. 2021 May 11;118(19):e2101918118.
doi: 10.1073/pnas.2101918118.
Nanobody cocktails potently neutralize SARS-CoV-2 D614G N501Y variant and protect mice
Phillip Pymm[SUP] 1 2 [/SUP], Amy Adair[SUP] 1 [/SUP], Li-Jin Chan[SUP] 1 2 [/SUP], James P Cooney[SUP] 1 2 [/SUP], Francesca L Mordant[SUP] 3 [/SUP], Cody C Allison[SUP] 1 2 [/SUP], Ester Lopez[SUP] 3 [/SUP], Ebene R Haycroft[SUP] 3 [/SUP], Matthew T O'Neill[SUP] 1 [/SUP], Li Lynn Tan[SUP] 1 [/SUP], Melanie H Dietrich[SUP] 1 2 [/SUP], Damien Drew[SUP] 1 [/SUP], Marcel Doerflinger[SUP] 1 2 [/SUP], Michael A Dengler[SUP] 1 2 [/SUP], Nichollas E Scott[SUP] 3 [/SUP], Adam K Wheatley[SUP] 3 4 [/SUP], Nicholas A Gherardin[SUP] 3 5 [/SUP], Hariprasad Venugopal[SUP] 6 [/SUP], Deborah Cromer[SUP] 7 8 [/SUP], Miles P Davenport[SUP] 7 [/SUP], Raelene Pickering[SUP] 9 [/SUP], Dale I Godfrey[SUP] 3 5 [/SUP], Damian F J Purcell[SUP] 3 [/SUP], Stephen J Kent[SUP] 3 4 [/SUP], Amy W Chung[SUP] 3 [/SUP], Kanta Subbarao[SUP] 3 10 [/SUP], Marc Pellegrini[SUP] 1 2 [/SUP], Alisa Glukhova[SUP] 1 11 12 [/SUP], Wai-Hong Tham[SUP] 13 2 [/SUP]
Affiliations
Abstract
Neutralizing antibodies are important for immunity against SARS-CoV-2 and as therapeutics for the prevention and treatment of COVID-19. Here, we identified high-affinity nanobodies from alpacas immunized with coronavirus spike and receptor-binding domains (RBD) that disrupted RBD engagement with the human receptor angiotensin-converting enzyme 2 (ACE2) and potently neutralized SARS-CoV-2. Epitope mapping, X-ray crystallography, and cryo-electron microscopy revealed two distinct antigenic sites and showed two neutralizing nanobodies from different epitope classes bound simultaneously to the spike trimer. Nanobody-Fc fusions of the four most potent nanobodies blocked ACE2 engagement with RBD variants present in human populations and potently neutralized both wild-type SARS-CoV-2 and the N501Y D614G variant at concentrations as low as 0.1 nM. Prophylactic administration of either single nanobody-Fc or as mixtures reduced viral loads by up to 10[SUP]4[/SUP]-fold in mice infected with the N501Y D614G SARS-CoV-2 virus. These results suggest a role for nanobody-Fc fusions as prophylactic agents against SARS-CoV-2.
Keywords: SARS-CoV-2; antiviral therapeutics; cryo-EM; crystallography; nanobodies.
. 2021 May 11;118(19):e2101918118.
doi: 10.1073/pnas.2101918118.
Nanobody cocktails potently neutralize SARS-CoV-2 D614G N501Y variant and protect mice
Phillip Pymm[SUP] 1 2 [/SUP], Amy Adair[SUP] 1 [/SUP], Li-Jin Chan[SUP] 1 2 [/SUP], James P Cooney[SUP] 1 2 [/SUP], Francesca L Mordant[SUP] 3 [/SUP], Cody C Allison[SUP] 1 2 [/SUP], Ester Lopez[SUP] 3 [/SUP], Ebene R Haycroft[SUP] 3 [/SUP], Matthew T O'Neill[SUP] 1 [/SUP], Li Lynn Tan[SUP] 1 [/SUP], Melanie H Dietrich[SUP] 1 2 [/SUP], Damien Drew[SUP] 1 [/SUP], Marcel Doerflinger[SUP] 1 2 [/SUP], Michael A Dengler[SUP] 1 2 [/SUP], Nichollas E Scott[SUP] 3 [/SUP], Adam K Wheatley[SUP] 3 4 [/SUP], Nicholas A Gherardin[SUP] 3 5 [/SUP], Hariprasad Venugopal[SUP] 6 [/SUP], Deborah Cromer[SUP] 7 8 [/SUP], Miles P Davenport[SUP] 7 [/SUP], Raelene Pickering[SUP] 9 [/SUP], Dale I Godfrey[SUP] 3 5 [/SUP], Damian F J Purcell[SUP] 3 [/SUP], Stephen J Kent[SUP] 3 4 [/SUP], Amy W Chung[SUP] 3 [/SUP], Kanta Subbarao[SUP] 3 10 [/SUP], Marc Pellegrini[SUP] 1 2 [/SUP], Alisa Glukhova[SUP] 1 11 12 [/SUP], Wai-Hong Tham[SUP] 13 2 [/SUP]
Affiliations
- PMID: 33893175
- DOI: 10.1073/pnas.2101918118
Abstract
Neutralizing antibodies are important for immunity against SARS-CoV-2 and as therapeutics for the prevention and treatment of COVID-19. Here, we identified high-affinity nanobodies from alpacas immunized with coronavirus spike and receptor-binding domains (RBD) that disrupted RBD engagement with the human receptor angiotensin-converting enzyme 2 (ACE2) and potently neutralized SARS-CoV-2. Epitope mapping, X-ray crystallography, and cryo-electron microscopy revealed two distinct antigenic sites and showed two neutralizing nanobodies from different epitope classes bound simultaneously to the spike trimer. Nanobody-Fc fusions of the four most potent nanobodies blocked ACE2 engagement with RBD variants present in human populations and potently neutralized both wild-type SARS-CoV-2 and the N501Y D614G variant at concentrations as low as 0.1 nM. Prophylactic administration of either single nanobody-Fc or as mixtures reduced viral loads by up to 10[SUP]4[/SUP]-fold in mice infected with the N501Y D614G SARS-CoV-2 virus. These results suggest a role for nanobody-Fc fusions as prophylactic agents against SARS-CoV-2.
Keywords: SARS-CoV-2; antiviral therapeutics; cryo-EM; crystallography; nanobodies.