Neutralizing nanobodies bind SARS-CoV-2 spike RBD and block interaction with ACE2

被引：16

作者：

Huo, Jiangdong ^{[1
,2
,3
]}

Le Bas, Audrey ^{[2
,3
]}

Ruza, Reinis R. ^{[2
]}

Duyvesteyn, Helen M. E. ^{[2
]}

Mikolajek, Halina ^{[4
]}

Malinauskas, Tomas ^{[2
]}

Tan, Tiong Kit ^{[5
]}

Rijal, Pramila ^{[5
,6
]}

Dumoux, Maud ^{[1
]}

Ward, Philip N. ^{[2
,3
]}

Ren, Jingshan ^{[2
]}

Zhou, Daming ^{[2
]}

Harrison, Peter J. ^{[2
,3
]}

Weckener, Miriam ^{[1
]}

Clare, Daniel K. ^{[4
]}

Vogirala, Vinod K. ^{[4
]}

Radecke, Julika ^{[4
]}

Moynie, Lucile ^{[1
]}

Zhao, Yuguang ^{[2
]}

Gilbert-Jaramillo, Javier ^{[7
]}

Knight, Michael L. ^{[7
]}

Tree, Julia A. ^{[8
]}

Buttigieg, Karen R. ^{[8
]}

Coombes, Naomi ^{[8
]}

Elmore, Michael J. ^{[8
]}

Carroll, Miles W. ^{[8
]}

Carrique, Loic ^{[2
]}

Shah, Pranav N. M. ^{[2
]}

James, William ^{[7
]}

Townsend, Alain R. ^{[5
,6
]}

Stuart, David, I ^{[2
,4
]}

Owens, Raymond J. ^{[1
,2
,3
]}

Naismith, James H. ^{[1
,2
,3
]}

机构：

[1] Harwell Sci & Innovat Campus, Rosalind Franklin Inst, Struct Biol, Didcot, Oxon, England

[2] Univ Oxford, Wellcome Ctr Human Genet, Div Struct Biol, Oxford, England

[3] Harwell Sci & Innovat Campus, Res Complex Harwell, Rosalind Franklin Inst Diamond Light Source, Prot Prod UK, Didcot, Oxon, England

[4] Harwell Sci & Innovat Campus, Diamond Light Source Ltd, Didcot, Oxon, England

[5] Univ Oxford, John Radcliffe Hosp, Weatherall Inst Mol Med, MRC Human Immunol Unit, Oxford, England

[6] Univ Oxford, Oxford Inst, Chinse Acad Med Sci, Ctr Translat Immunol, Oxford, England

[7] Univ Oxford, Sir William Dunn Sch Pathol, Oxford, England

[8] Publ Hlth England, Natl Infect Serv, Salisbury, Wilts, England

来源：

NATURE STRUCTURAL & MOLECULAR BIOLOGY | 2020年 / 27卷 / 09期

基金：

英国惠康基金; 英国生物技术与生命科学研究理事会; 英国医学研究理事会; 英国工程与自然科学研究理事会;

关键词：

CRYO-EM STRUCTURE; PROTEIN; REFINEMENT; REFMAC5; SYSTEM;

D O I：

10.1038/s41594-020-0469-6

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Two nanobodies that bind SARS-CoV-2 spike RBD are shown to block interaction with receptor ACE2 and thus neutralize the virus, and have an additive effect with antibody CR3022. 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(D)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.

引用

页码：846 / +

页数：25

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