A Potently Neutralizing Antibody Protects Mice against SARS-CoV-2 Infection

Key Points Generation and screening of anti–SARS-CoV-2 RBD mAbs reveal a potently protective Ab. mAb 2B04 potently neutralizes SARS-CoV-2 in vitro. mAb 2B04 protects mice from morbidity in a murine model of SARS-CoV-2 infection. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for millions of infections and hundreds of thousands of deaths globally. There are no widely available licensed therapeutics against SARS-CoV-2, highlighting an urgent need for effective interventions. The virus enters host cells through binding of a receptor-binding domain within its trimeric spike glycoprotein to human angiotensin-converting enzyme 2. In this article, we describe the generation and characterization of a panel of murine mAbs directed against the receptor-binding domain. One mAb, 2B04, neutralized wild-type SARS-CoV-2 in vitro with remarkable potency (half-maximal inhibitory concentration of <2 ng/ml). In a murine model of SARS-CoV-2 infection, 2B04 protected challenged animals from weight loss, reduced lung viral load, and blocked systemic dissemination. Thus, 2B04 is a promising candidate for an effective antiviral that can be used to prevent SARS-CoV-2 infection.

[1]  Larissa B. Thackray,et al.  A SARS-CoV-2 Infection Model in Mice Demonstrates Protection by Neutralizing Antibodies , 2020, Cell.

[2]  Linqi Zhang,et al.  Human neutralizing antibodies elicited by SARS-CoV-2 infection , 2020, Nature.

[3]  William J. Liu,et al.  A human neutralizing antibody targets the receptor-binding site of SARS-CoV-2 , 2020, Nature.

[4]  L. Guddat,et al.  Structural Basis for RNA Replication by the SARS-CoV-2 Polymerase , 2020, Cell.

[5]  X. Xie,et al.  Potent Neutralizing Antibodies against SARS-CoV-2 Identified by High-Throughput Single-Cell Sequencing of Convalescent Patients’ B Cells , 2020, Cell.

[6]  H. Yen,et al.  Peer Review File Manuscript Title: Pathogenesis and transmission of SARS-CoV-2 in golden Syrian hamsters , 2020 .

[7]  A. Iwasaki,et al.  The potential danger of suboptimal antibody responses in COVID-19 , 2020, Nature Reviews Immunology.

[8]  K. Yuen,et al.  Structural and Functional Basis of SARS-CoV-2 Entry by Using Human ACE2 , 2020, Cell.

[9]  I. Wilson,et al.  A highly conserved cryptic epitope in the receptor binding domains of SARS-CoV-2 and SARS-CoV , 2020, Science.

[10]  L. Poon,et al.  Pathogenesis and transmission of SARS-CoV-2 virus in golden Syrian hamsters , 2020 .

[11]  K. Shi,et al.  Structural basis of receptor recognition by SARS-CoV-2 , 2020, Nature.

[12]  Shibo Jiang,et al.  Characterization of the receptor-binding domain (RBD) of 2019 novel coronavirus: implication for development of RBD protein as a viral attachment inhibitor and vaccine , 2020, Cellular & Molecular Immunology.

[13]  Nicholas C. Wu,et al.  A highly conserved cryptic epitope in the receptor binding domains of SARS-CoV-2 and SARS-CoV , 2020, Science.

[14]  Frank Grosveld,et al.  A human monoclonal antibody blocking SARS-CoV-2 infection , 2020, Nature Communications.

[15]  Vineet D. Menachery,et al.  Severe Acute Respiratory Syndrome Coronavirus 2 from Patient with Coronavirus Disease, United States , 2020, Emerging infectious diseases.

[16]  Kai Zhao,et al.  A pneumonia outbreak associated with a new coronavirus of probable bat origin , 2020, Nature.

[17]  Jing Zhao,et al.  Early Transmission Dynamics in Wuhan, China, of Novel Coronavirus–Infected Pneumonia , 2020, The New England journal of medicine.

[18]  G. Gao,et al.  A Novel Coronavirus from Patients with Pneumonia in China, 2019 , 2020, The New England journal of medicine.

[19]  S. Lo,et al.  A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: a study of a family cluster , 2020, The Lancet.

[20]  Y. Hu,et al.  Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China , 2020, The Lancet.

[21]  Astrid Gall,et al.  Ensembl 2020 , 2019, Nucleic Acids Res..

[22]  C. E..  WHO Coronavirus Disease (COVID-19) Dashboard , 2020 .

[23]  M. Levine.  Monoclonal Antibody Therapy for Ebola Virus Disease. , 2019, The New England journal of medicine.

[24]  V. Corman,et al.  [Coronaviruses as the cause of respiratory infections]. , 2019, Der Internist.

[25]  Paul J. Hoffman,et al.  Comprehensive Integration of Single-Cell Data , 2018, Cell.

[26]  J. Sidney,et al.  Longitudinal Analysis of the Human B Cell Response to Ebola Virus Infection , 2019, Cell.

[27]  Linqi Zhang,et al.  Antibodies and vaccines against Middle East respiratory syndrome coronavirus , 2019, Emerging microbes & infections.

[28]  Judith A. Blake,et al.  Mouse Genome Database (MGD) 2019 , 2018, Nucleic Acids Res..

[29]  Francesco Vallania,et al.  Leveraging heterogeneity across multiple datasets increases cell-mixture deconvolution accuracy and reduces biological and technical biases , 2018, Nature Communications.

[30]  Christoph Hafemeister,et al.  Comprehensive integration of single cell data , 2018, bioRxiv.

[31]  Catharine I Paules,et al.  Monoclonal Antibodies for Emerging Infectious Diseases - Borrowing from History. , 2018, The New England journal of medicine.

[32]  Barney S. Graham,et al.  Immunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen , 2017, Proceedings of the National Academy of Sciences.

[33]  Francois Vigneault,et al.  Hierarchical Clustering Can Identify B Cell Clones with High Confidence in Ig Repertoire Sequencing Data , 2017, The Journal of Immunology.

[34]  Karlynn E. Neu,et al.  Refined protocol for generating monoclonal antibodies from single human and murine B cells. , 2016, Journal of immunological methods.

[35]  Fang Li,et al.  Structure, Function, and Evolution of Coronavirus Spike Proteins. , 2016, Annual review of virology.

[36]  M. Nussenzweig,et al.  Sequencing and cloning of antigen-specific antibodies from mouse memory B cells , 2016, Nature Protocols.

[37]  C. Nelson,et al.  Structural Basis of Zika Virus-Specific Antibody Protection , 2016, Cell.

[38]  J. Mascola,et al.  Protective monotherapy against lethal Ebola virus infection by a potently neutralizing antibody , 2016, Science.

[39]  Steven H. Kleinstein,et al.  Change-O: a toolkit for analyzing large-scale B cell immunoglobulin repertoire sequencing data , 2015, Bioinform..

[40]  Alessandro Vullo,et al.  The Ensembl REST API: Ensembl Data for Any Language , 2014, Bioinform..

[41]  David A. Hafler,et al.  pRESTO: a toolkit for processing high-throughput sequencing raw reads of lymphocyte receptor repertoires , 2014, Bioinform..

[42]  N. Neff,et al.  Reconstructing lineage hierarchies of the distal lung epithelium using single cell RNA-seq , 2014, Nature.

[43]  Ning Ma,et al.  IgBLAST: an immunoglobulin variable domain sequence analysis tool , 2013, Nucleic Acids Res..

[44]  A. Osterhaus,et al.  Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. , 2012, The New England journal of medicine.

[45]  R. Wilson,et al.  Modernizing Reference Genome Assemblies , 2011, PLoS biology.

[46]  T. Tiller,et al.  Cloning and expression of murine Ig genes from single B cells. , 2009, Journal of immunological methods.

[47]  Shibo Jiang,et al.  The spike protein of SARS-CoV — a target for vaccine and therapeutic development , 2009, Nature Reviews Microbiology.

[48]  M. Ernstoff,et al.  The clinical pharmacology of therapeutic monoclonal antibodies in the treatment of malignancy; have the magic bullets arrived? , 2008, British journal of clinical pharmacology.

[49]  K. P. Murphy,et al.  Janeway's immunobiology , 2007 .

[50]  Y. Nishida,et al.  Housekeeping and tissue-specific genes in mouse tissues , 2007, BMC Genomics.

[51]  R. Schreiber,et al.  Blocking monoclonal antibodies specific for mouse IFN-alpha/beta receptor subunit 1 (IFNAR-1) from mice immunized by in vivo hydrodynamic transfection. , 2006, Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research.

[52]  A. Aderem,et al.  TLR9/MyD88 signaling is required for class switching to pathogenic IgG2a and 2b autoantibodies in SLE , 2006, The Journal of Experimental Medicine.

[53]  S. Perlman,et al.  ACE2 Receptor Expression and Severe Acute Respiratory Syndrome Coronavirus Infection Depend on Differentiation of Human Airway Epithelia , 2005, Journal of Virology.

[54]  Kwanyee Leung,et al.  Evasion of antibody neutralization in emerging severe acute respiratory syndrome coronaviruses. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[55]  Marie-Paule Lefranc,et al.  IMGT , the international ImMunoGeneTics information system , 2003 .

[56]  Antony V. Cox,et al.  The Ensembl Web site: mechanics of a genome browser. , 2004, Genome research.

[57]  B. Bosch,et al.  The Coronavirus Spike Protein Is a Class I Virus Fusion Protein: Structural and Functional Characterization of the Fusion Core Complex , 2003, Journal of Virology.

[58]  Christian Drosten,et al.  Identification of a novel coronavirus in patients with severe acute respiratory syndrome. , 2003, The New England journal of medicine.

[59]  J. A. Comer,et al.  A novel coronavirus associated with severe acute respiratory syndrome. , 2003, The New England journal of medicine.

[60]  V. Giudicelli,et al.  IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains. , 2003, Developmental and comparative immunology.

[61]  B. Trapnell,et al.  Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy , 1996, Journal of virology.

[62]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.