"Isolation, characterization, and structure-based engineering of a neutralizing nanobody against SARS-CoV-2"

作者全名："Li, Tingting; Zhou, Bingjie; Li, Yaning; Huang, Suqiong; Luo, Zhipu; Zhou, Yuanze; Lai, Yanling; Gautam, Anupriya; Bourgeau, Salome; Wang, Shurui; Bao, Juan; Tan, Jingquan; Lavillette, Dimitri; Li, Dianfan"

作者地址："[Li, Tingting; Li, Yaning; Lai, Yanling; Bao, Juan; Li, Dianfan] Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, CAS Ctr Excellence Mol Cell Sci, State Key Lab Mol Biol, 320 Yueyang Rd, Shanghai 200030, Peoples R China; [Zhou, Bingjie; Li, Yaning; Huang, Suqiong; Lai, Yanling; Gautam, Anupriya; Bourgeau, Salome] Univ CAS, Beijing 101408, Peoples R China; [Zhou, Bingjie; Huang, Suqiong; Gautam, Anupriya; Bourgeau, Salome; Lavillette, Dimitri] Inst Pasteur Shanghai CAS, CAS Key Lab Mol Virol & Immunol, 320 Yueyang Rd, Shanghai 200031, Peoples R China; [Huang, Suqiong] Chongqing Med Univ, Coll Pharm, Chongqing, Peoples R China; [Luo, Zhipu] Soochow Univ, Inst Mol Enzymol, Sch Biol & Basic Med Sci, Suzhou, Peoples R China; [Zhou, Yuanze; Wang, Shurui; Tan, Jingquan] Nanjing Crycis Biotech Co Ltd, Nanjing, Peoples R China; [Bourgeau, Salome] Univ Rennes, Ecole Hautes Etud Sante Publ, Inst Natl Sante & Rech Med, Inst Rech Sante Environm & Travail, F-35000 Rennes, France; [Lavillette, Dimitri] Soochow Univ, Pasteurien Coll, Suzhou, Jiangsu, Peoples R China"

通信作者："Li, DF (通讯作者)，Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol, CAS Ctr Excellence Mol Cell Sci, State Key Lab Mol Biol, 320 Yueyang Rd, Shanghai 200030, Peoples R China.; Lavillette, D (通讯作者)，Inst Pasteur Shanghai CAS, CAS Key Lab Mol Virol & Immunol, 320 Yueyang Rd, Shanghai 200031, Peoples R China.; Lavillette, D (通讯作者)，Soochow Univ, Pasteurien Coll, Suzhou, Jiangsu, Peoples R China."

来源：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:000803813700008

JCR分区：Q1

影响因子：8.2

年份：2022

卷号：209

期号：　

开始页：1379

结束页：1388

文献类型：Article

关键词：Covid-19; Crystal structure; Neutralizing antibody; Protein engineering; Receptor-binding domain; Single-chain antibody

摘要："SARS-CoV-2 engages with human cells through the binding of its Spike receptor-binding domain (S-RBD) to the receptor ACE2. Molecular blocking of this engagement represents a proven strategy to treat COVID-19. Here, we report a single-chain antibody (nanobody, DL4) isolated from immunized alpaca with picomolar affinity to RBD. DL4 neutralizes SARS-CoV-2 pseudoviruses with an IC50 of 0.101 mu g mL-1 (6.2 nM). A crystal structure of the DL4-RBD complex at 1.75-angstrom resolution unveils the interaction detail and reveals a direct competition mechanism for DL4's ACE2-blocking and hence neutralizing activity. The structural information allows us to rationally design a mutant with higher potency. Our work adds diversity of neutralizing nanobodies against SARS-CoV-2 and should encourage protein engineering to improve antibody affinities in general."

基金机构："National Natural Science Foundation of China [82151215, 31870726, 31870153]; Strategic Priority Research Program of CAS [XDB37020204]; Key Program of CAS Frontier Science [QYZDB-SSW-SMC037]; CAS Facility-based Open Research Program [2020YFC0845900]; National Key Research and Development Program of China [2020VBA0023]; CAS president's international fellowship initiative [20ZR1466700]; Science and Technology Commission of Shanghai Municipality [20431900402]; Shanghai~Municipal Science and Technology Major Project"

基金资助正文："We thank the staff scientists at the SSRF-BL18U1 beamline at National Facility for Protein Science (Shanghai) for technical support, and Dr. Peiling Li from Xinyang Agricultural and Forestry University for assistance in the alpaca immunization. This work has been supported by the National Natural Science Foundation of China (82151215, 31870726, D. Li; 31870153, D. La), the Strategic Priority Research Program of CAS (XDB37020204, D. Li), Key Program of CAS Frontier Science (QYZDB-SSW-SMC037, D. Li), CAS Facility-based Open Research Program, National Key Research and Development Program of China (2020YFC0845900, D. La), CAS president's international fellow-ship initiative (2020VBA0023, D. La), Science and Technology Commission of Shanghai Municipality (20ZR1466700, D. Li), Shanghai Municipal Science and Technology Major Project (20431900402, D. La)"