An alpaca nanobody neutralizes SARS-CoV-2 by blocking receptor interaction

被引:251
作者
Hanke, Leo [1 ]
Perez, Laura Vidakovics [1 ]
Sheward, Daniel J. [1 ,2 ]
Das, Hrishikesh [3 ]
Schulte, Tim [4 ,5 ]
Moliner-Morro, Ainhoa [1 ]
Corcoran, Martin [1 ]
Achour, Adnane [4 ,5 ]
Hedestam, Gunilla B. Karlsson [1 ]
Haellberg, B. Martin [3 ,6 ]
Murrell, Ben [1 ]
McInerney, Gerald M. [1 ]
机构
[1] Karolinska Inst, Dept Microbiol Tumor & Cell Biol, Stockholm, Sweden
[2] Univ Cape Town, Fac Hlth Sci, Inst Infect Dis & Mol Med, Div Virol, Cape Town, South Africa
[3] Karolinska Inst, Dept Cell & Mol Biol, Stockholm, Sweden
[4] Karolinska Inst, Dept Med Solna, Sci Life Lab, Stockholm, Sweden
[5] Karolinska Univ Hosp, Div Infect Dis, Stockholm, Sweden
[6] Karolinska Inst, VR RAC, Ctr Struct Syst Biol, Notkestr 85, D-22607 Hamburg, Germany
基金
欧盟地平线“2020”; 瑞典研究理事会;
关键词
SINGLE-DOMAIN ANTIBODIES; CRYO-EM STRUCTURE; PROTEIN INTERACTIONS; POTENT; HETEROGENEITY; GENERATION; BIOSENSOR; KINETICS; AFFINITY;
D O I
10.1038/s41467-020-18174-5
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
SARS-CoV-2 enters host cells through an interaction between the spike glycoprotein and the angiotensin converting enzyme 2 (ACE2) receptor. Directly preventing this interaction presents an attractive possibility for suppressing SARS-CoV-2 replication. Here, we report the isolation and characterization of an alpaca-derived single domain antibody fragment, Ty1, that specifically targets the receptor binding domain (RBD) of the SARS-CoV-2 spike, directly preventing ACE2 engagement. Ty1 binds the RBD with high affinity, occluding ACE2. A cryo-electron microscopy structure of the bound complex at 2.9 angstrom resolution reveals that Ty1 binds to an epitope on the RBD accessible in both the 'up' and 'down' conformations, sterically hindering RBD-ACE2 binding. While fusion to an Fc domain renders Ty1 extremely potent, Ty1 neutralizes SARS-CoV-2 spike pseudovirus as a 12.8kDa nanobody, which can be expressed in high quantities in bacteria, presenting opportunities for manufacturing at scale. Ty1 is therefore an excellent candidate as an intervention against COVID-19. Here, Hanke et al. immunize an alpaca with SARS-CoV-2 spike protein domains and identify a nanobody that binds the receptor binding domain of spike in both the up and down conformations and sterically hinders ACE2 engagement.
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页数:9
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