TY - JOUR
T1 - A synthetic nanobody targeting RBD protects hamsters from SARS-CoV-2 infection
AU - Li, Tingting
AU - Cai, Hongmin
AU - Yao, Hebang
AU - Zhou, Bingjie
AU - Zhang, Ning
AU - van Vlissingen, Martje Fentener
AU - Kuiken, Thijs
AU - Han, Wenyu
AU - GeurtsvanKessel, Corine H.
AU - Gong, Yuhuan
AU - Zhao, Yapei
AU - Shen, Quan
AU - Qin, Wenming
AU - Tian, Xiao Xu
AU - Peng, Chao
AU - Lai, Yanling
AU - Wang, Yanxing
AU - Hutter, Cedric A.J.
AU - Kuo, Shu Ming
AU - Bao, Juan
AU - Liu, Caixuan
AU - Wang, Yifan
AU - Richard, Audrey S.
AU - Raoul, Hervé
AU - Lan, Jiaming
AU - Seeger, Markus A.
AU - Cong, Yao
AU - Rockx, Barry
AU - Wong, Gary
AU - Bi, Yuhai
AU - Lavillette, Dimitri
AU - Li, Dianfan
N1 - Funding Information:
We thank the staff members of the NCPSS Large-scale Protein Preparation System and the Electron Microscopy facility for equipment maintenance and management and staff scientists at the SSRF-BL19U1 beamline at National Facility for Protein Science (Shanghai), and the staff of BSL-3 in the Institute of Microbiology, Chinese Academy of Sciences for technical support and assistance. We thank Dr. Zhipu Luo at Soochow University (China) for helpful discussions regarding data processing. This work has been supported by the Strategic Priority Research Program of CAS (XDB37020204, D. Li; XDB29010102, Y.B.), Key Program of CAS Frontier Science (QYZDB-SSW-SMC037, D. Li), CAS Facility-based Open Research Program, the National Natural Science Foundation of China (31870726, D. Li; 31870153, D. Lavillette; 32041010, Y.B.), the One Belt and One Road major project for infectious diseases (2018ZX10101004-003, J.L., G.W.), National Key R&D Program of China (2020YFC0845900, D. Lavillette), CAS president’s international fellowship initiative (2020VBA0023, D. Lavillette), Natural Science Foundation of Shanghai (20ZR1466700, D. Li; 20ZR1463900, G.W.), Shanghai Municipal Science and Technology Major Project (20431900402, D. Lavillette), Science and Technology Commission of Shanghai Municipality (18DZ2210200), Funding for Construction and Operation of Zhangjiang Laboratory (II) (19DZ2260100), and the ERINHA-Advance project (funding from the European Union’s Horizon 2020 Research & Innovation program, grant agreement No. 824061). This project is included in RECOVER European Union’s Horizon 2020 research and innovation program under grant agreement No. 101003589. Y.B. is supported by the NSFC Outstanding Young Scholars (31822055) and Youth Innovation Promotion Association of CAS (2017122). G.W. is supported by a G4 grant from IP, FMX, and CAS.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/7/30
Y1 - 2021/7/30
N2 - SARS-CoV-2, the causative agent of COVID-191, features a receptor-binding domain (RBD) for binding to the host cell ACE2 protein1–6. Neutralizing antibodies that block RBD-ACE2 interaction are candidates for the development of targeted therapeutics7–17. Llama-derived single-domain antibodies (nanobodies, ~15 kDa) offer advantages in bioavailability, amenability, and production and storage owing to their small sizes and high stability. Here, we report the rapid selection of 99 synthetic nanobodies (sybodies) against RBD by in vitro selection using three libraries. The best sybody, MR3 binds to RBD with high affinity (KD = 1.0 nM) and displays high neutralization activity against SARS-CoV-2 pseudoviruses (IC50 = 0.42 μg mL−1). Structural, biochemical, and biological characterization suggests a common neutralizing mechanism, in which the RBD-ACE2 interaction is competitively inhibited by sybodies. Various forms of sybodies with improved potency have been generated by structure-based design, biparatopic construction, and divalent engineering. Two divalent forms of MR3 protect hamsters from clinical signs after live virus challenge and a single dose of the Fc-fusion construct of MR3 reduces viral RNA load by 6 Log10. Our results pave the way for the development of therapeutic nanobodies against COVID-19 and present a strategy for rapid development of targeted medical interventions during an outbreak.
AB - SARS-CoV-2, the causative agent of COVID-191, features a receptor-binding domain (RBD) for binding to the host cell ACE2 protein1–6. Neutralizing antibodies that block RBD-ACE2 interaction are candidates for the development of targeted therapeutics7–17. Llama-derived single-domain antibodies (nanobodies, ~15 kDa) offer advantages in bioavailability, amenability, and production and storage owing to their small sizes and high stability. Here, we report the rapid selection of 99 synthetic nanobodies (sybodies) against RBD by in vitro selection using three libraries. The best sybody, MR3 binds to RBD with high affinity (KD = 1.0 nM) and displays high neutralization activity against SARS-CoV-2 pseudoviruses (IC50 = 0.42 μg mL−1). Structural, biochemical, and biological characterization suggests a common neutralizing mechanism, in which the RBD-ACE2 interaction is competitively inhibited by sybodies. Various forms of sybodies with improved potency have been generated by structure-based design, biparatopic construction, and divalent engineering. Two divalent forms of MR3 protect hamsters from clinical signs after live virus challenge and a single dose of the Fc-fusion construct of MR3 reduces viral RNA load by 6 Log10. Our results pave the way for the development of therapeutic nanobodies against COVID-19 and present a strategy for rapid development of targeted medical interventions during an outbreak.
UR - http://www.scopus.com/inward/record.url?scp=85111631022&partnerID=8YFLogxK
U2 - 10.1038/s41467-021-24905-z
DO - 10.1038/s41467-021-24905-z
M3 - Article
AN - SCOPUS:85111631022
VL - 12
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 4635
ER -