tetano
Editor, Senior Moderator
Nat Struct Mol Biol
. 2020 Jul 13.
doi: 10.1038/s41594-020-0469-6. Online ahead of print.
Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2
Jiangdong Huo[SUP] 1 2 3 [/SUP], Audrey Le Bas[SUP] 2 3 [/SUP], Reinis R Ruza[SUP] 2 [/SUP], Helen M E Duyvesteyn[SUP] 2 [/SUP], Halina Mikolajek[SUP] 4 [/SUP], Tomas Malinauskas[SUP] 2 [/SUP], Tiong Kit Tan[SUP] 5 [/SUP], Pramila Rijal[SUP] 5 6 [/SUP], Maud Dumoux[SUP] 1 [/SUP], Philip N Ward[SUP] 2 3 [/SUP], Jingshan Ren[SUP] 2 [/SUP], Daming Zhou[SUP] 2 [/SUP], Peter J Harrison[SUP] 2 3 [/SUP], Miriam Weckener[SUP] 1 [/SUP], Daniel K Clare[SUP] 4 [/SUP], Vinod K Vogirala[SUP] 4 [/SUP], Julika Radecke[SUP] 4 [/SUP], Lucile Moyni?[SUP] 1 [/SUP], Yuguang Zhao[SUP] 2 [/SUP], Javier Gilbert-Jaramillo[SUP] 7 [/SUP], Michael L Knight[SUP] 7 [/SUP], Julia A Tree[SUP] 8 [/SUP], Karen R Buttigieg[SUP] 8 [/SUP], Naomi Coombes[SUP] 8 [/SUP], Michael J Elmore[SUP] 8 [/SUP], Miles W Carroll[SUP] 8 [/SUP], Loic Carrique[SUP] 2 [/SUP], Pranav N M Shah[SUP] 2 [/SUP], William James[SUP] 7 [/SUP], Alain R Townsend[SUP] 5 6 [/SUP], David I Stuart[SUP] 2 4 [/SUP], Raymond J Owens[SUP] 9 10 11 [/SUP], James H Naismith[SUP] 12 13 14 [/SUP]
Affiliations
Abstract
The SARS-CoV-2 virus is more transmissible than previous coronaviruses and causes a more serious illness than influenza. The SARS-CoV-2 receptor binding domain (RBD) of the spike protein binds to the human angiotensin-converting enzyme 2 (ACE2) receptor as a prelude to viral entry into the cell. Using a naive llama single-domain antibody library and PCR-based maturation, we have produced two closely related nanobodies, H11-D4 and H11-H4, that bind RBD (K[SUB]D[/SUB] of 39 and 12 nM, respectively) and block its interaction with ACE2. Single-particle cryo-EM revealed that both nanobodies bind to all three RBDs in the spike trimer. Crystal structures of each nanobody-RBD complex revealed how both nanobodies recognize the same epitope, which partly overlaps with the ACE2 binding surface, explaining the blocking of the RBD-ACE2 interaction. Nanobody-Fc fusions showed neutralizing activity against SARS-CoV-2 (4-6 nM for H11-H4, 18 nM for H11-D4) and additive neutralization with the SARS-CoV-1/2 antibody CR3022.
. 2020 Jul 13.
doi: 10.1038/s41594-020-0469-6. Online ahead of print.
Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2
Jiangdong Huo[SUP] 1 2 3 [/SUP], Audrey Le Bas[SUP] 2 3 [/SUP], Reinis R Ruza[SUP] 2 [/SUP], Helen M E Duyvesteyn[SUP] 2 [/SUP], Halina Mikolajek[SUP] 4 [/SUP], Tomas Malinauskas[SUP] 2 [/SUP], Tiong Kit Tan[SUP] 5 [/SUP], Pramila Rijal[SUP] 5 6 [/SUP], Maud Dumoux[SUP] 1 [/SUP], Philip N Ward[SUP] 2 3 [/SUP], Jingshan Ren[SUP] 2 [/SUP], Daming Zhou[SUP] 2 [/SUP], Peter J Harrison[SUP] 2 3 [/SUP], Miriam Weckener[SUP] 1 [/SUP], Daniel K Clare[SUP] 4 [/SUP], Vinod K Vogirala[SUP] 4 [/SUP], Julika Radecke[SUP] 4 [/SUP], Lucile Moyni?[SUP] 1 [/SUP], Yuguang Zhao[SUP] 2 [/SUP], Javier Gilbert-Jaramillo[SUP] 7 [/SUP], Michael L Knight[SUP] 7 [/SUP], Julia A Tree[SUP] 8 [/SUP], Karen R Buttigieg[SUP] 8 [/SUP], Naomi Coombes[SUP] 8 [/SUP], Michael J Elmore[SUP] 8 [/SUP], Miles W Carroll[SUP] 8 [/SUP], Loic Carrique[SUP] 2 [/SUP], Pranav N M Shah[SUP] 2 [/SUP], William James[SUP] 7 [/SUP], Alain R Townsend[SUP] 5 6 [/SUP], David I Stuart[SUP] 2 4 [/SUP], Raymond J Owens[SUP] 9 10 11 [/SUP], James H Naismith[SUP] 12 13 14 [/SUP]
Affiliations
- PMID: 32661423
- DOI: 10.1038/s41594-020-0469-6
Abstract
The SARS-CoV-2 virus is more transmissible than previous coronaviruses and causes a more serious illness than influenza. The SARS-CoV-2 receptor binding domain (RBD) of the spike protein binds to the human angiotensin-converting enzyme 2 (ACE2) receptor as a prelude to viral entry into the cell. Using a naive llama single-domain antibody library and PCR-based maturation, we have produced two closely related nanobodies, H11-D4 and H11-H4, that bind RBD (K[SUB]D[/SUB] of 39 and 12 nM, respectively) and block its interaction with ACE2. Single-particle cryo-EM revealed that both nanobodies bind to all three RBDs in the spike trimer. Crystal structures of each nanobody-RBD complex revealed how both nanobodies recognize the same epitope, which partly overlaps with the ACE2 binding surface, explaining the blocking of the RBD-ACE2 interaction. Nanobody-Fc fusions showed neutralizing activity against SARS-CoV-2 (4-6 nM for H11-H4, 18 nM for H11-D4) and additive neutralization with the SARS-CoV-1/2 antibody CR3022.