机构：[1]CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Collaborative Innovation Center forDiagnosis and Treatment of Infectious Diseases, Chinese Academy of Sciences, Beijing 100101, China[2]School of Life Sciences,Anhui University, Hefei, Anhui 230039, China[3]CAS Key Laboratory of Microbial and Metabolic engineering, Institute of Microbiology,Chinese Academy of Sciences, Beijing 100101, China[4]Departments of Microbiology and Pediatrics, University of Iowa,Iowa City, IA 52242, USA[5]State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China MedicalSchool, Sichuan University, Chengdu, Sichuan 610041, China四川大学华西医院[6]Laboratory Animal Center, Chinese Center for Disease Controland Prevention (China CDC), Beijing 102206, China[7]State Key Laboratory for Emerging Infectious Diseases, Department ofMicrobiology, The University of Hong Kong, Pokfulam, Hong Kong SAR, China[8]National Institute for Viral Disease Control andPrevention, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206, China[9]China-Japan Joint Laboratoryof Molecular Microbiology and Molecular Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing100101, China[10]Office of Director-General, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206,China[11]State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospitalof Guangzhou Medical University, Guangzhou, Guangdong 510120, China
The newly-emerging Middle East respiratory syndrome coronavirus (MERS-CoV) can cause severe and fatal acute respiratory disease in humans. Despite global efforts, the potential for an associated pandemic in the future cannot be excluded. The development of effective counter-measures is urgent. MERS-CoV-specific anti-viral drugs or vaccines are not yet available. Using the spike receptor-binding domain of MERS-CoV (MERS-RBD) to immunize mice, we identified two neutralizing monoclonal antibodies (mAbs) 4C2 and 2E6. Both mAbs potently bind to MERS-RBD and block virus entry in vitro with high efficacy. We further investigated their mechanisms of neutralization by crystallizing the complex between the Fab fragments and the RBD, and solved the structure of the 4C2 Fab/MERS-RBD complex. The structure showed that 4C2 recognizes an epitope that partially overlaps the receptor-binding footprint in MERS-RBD, thereby interfering with the virus/receptor interactions by both steric hindrance and interface-residue competition. 2E6 also blocks receptor binding, and competes with 4C2 for binding to MERS-RBD. Based on the structure, we further humanized 4C2 by preserving only the paratope residues and substituting the remaining amino acids with the counterparts from human immunoglobulins. The humanized 4C2 (4C2h) antibody sustained similar neutralizing activity and biochemical characteristics to the parental mouse antibody. Finally, we showed that 4C2h can significantly abate the virus titers in lungs of Ad5-hCD26-transduced mice infected with MERS-CoV, therefore representing a promising agent for prophylaxis and therapy in clinical settings.
基金：
the National Basic Research Program
of China (973 Program; 2015CB910503), the National Science
and Technology Major Project of China (2013ZX10004608-
002 and 2014ZX10004001-006), the National Natural Science
Foundation of China (NSFC; 31400154 and 81461168030), the
National Institutes of Health, USA (AI60699) and Hong Kong
Research Grants Council (NHKU728/14). We acknowledge Dr
Kwok-Hung Chan and Dr Jasper FW Chan for their technical support
and the staff at the Shanghai Synchrotron Radiation Facility
(SSRF-beam line 17U). We would be grateful to Yuanyuan Chen
(Institute of Biophysics, CAS) for technical help with Biacore
experiments. GFG is a leading principal investigator of the NSFC
Innovative Research Group (8132106).
语种：
外文
PubmedID：
中科院(CAS)分区：
出版当年[2015]版：
大类|1 区生物
小类|2 区细胞生物学
最新[2023]版：
大类|1 区生物学
小类|1 区细胞生物学
第一作者：
第一作者机构：[1]CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Collaborative Innovation Center forDiagnosis and Treatment of Infectious Diseases, Chinese Academy of Sciences, Beijing 100101, China
共同第一作者：
通讯作者：
通讯机构：[1]CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Collaborative Innovation Center forDiagnosis and Treatment of Infectious Diseases, Chinese Academy of Sciences, Beijing 100101, China[2]School of Life Sciences,Anhui University, Hefei, Anhui 230039, China[3]CAS Key Laboratory of Microbial and Metabolic engineering, Institute of Microbiology,Chinese Academy of Sciences, Beijing 100101, China[9]China-Japan Joint Laboratoryof Molecular Microbiology and Molecular Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing100101, China[10]Office of Director-General, Chinese Center for Disease Control and Prevention (China CDC), Beijing 102206,China[11]State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospitalof Guangzhou Medical University, Guangzhou, Guangdong 510120, China
推荐引用方式(GB/T 7714)：
Li Yan,Wan Yuhua,Liu Peipei,et al.A humanized neutralizing antibody against MERS-CoV targeting the receptor-binding domain of the spike protein.[J].Cell Research.2015,25(11):1237-49.doi:10.1038/cr.2015.113.
APA：
Li Yan,Wan Yuhua,Liu Peipei,Zhao Jincun,Lu Guangwen...&Yan Jinghua.(2015).A humanized neutralizing antibody against MERS-CoV targeting the receptor-binding domain of the spike protein..Cell Research,25,(11)
MLA：
Li Yan,et al."A humanized neutralizing antibody against MERS-CoV targeting the receptor-binding domain of the spike protein.".Cell Research 25..11(2015):1237-49
生物