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Identification of neutralizing human monoclonal antibodies from Italian Covid-19 convalescent patients
Emanuele Andreano; Emanuele Nicastri; Ida Paciello; Piero Pileri; Noemi Manganaro; Giulia Piccini; Alessandro Manenti; Marco Troisi; Fabiola Vacca; Elisa Pantano; Concetta De Santi; Dario Cardamone; Anna Kabanova; Chiara Agrati; Concetta Castilletti; Maria Rosaria Capobianchi; Arianna Emiliozzi; Massimiliano Fabbiani; Francesca Montagnani; Emanuele Montomoli; Claudia Sala; Giuseppe Ippolito; Rino Rappuoli.
Affiliation
  • Emanuele Andreano; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Emanuele Nicastri; National Institute for Infectious Diseases Lazzaro Spallanzani, IRCCS, Rome, Italy
  • Ida Paciello; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Piero Pileri; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Noemi Manganaro; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Giulia Piccini; VisMederi S.r.l, Siena, Italy
  • Alessandro Manenti; VisMederi S.r.l, Siena, Italy; VisMederi Research S.r.l., Siena, Italy
  • Marco Troisi; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Fabiola Vacca; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Elisa Pantano; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Concetta De Santi; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Dario Cardamone; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Anna Kabanova; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy; Tumour Immunology Unit, Fondazione Toscana Life Sciences, Siena, Italy
  • Chiara Agrati; National Institute for Infectious Diseases Lazzaro Spallanzani, IRCCS, Rome, Italy
  • Concetta Castilletti; National Institute for Infectious Diseases Lazzaro Spallanzani, IRCCS, Rome, Italy
  • Maria Rosaria Capobianchi; National Institute for Infectious Diseases Lazzaro Spallanzani, IRCCS, Rome, Italy
  • Arianna Emiliozzi; Department of Medical Biotechnologies, University of Siena, Siena, Italy; Department of Specialized and Internal Medicine, Tropical and Infectious Diseases Unit
  • Massimiliano Fabbiani; Department of Specialized and Internal Medicine, Tropical and Infectious Diseases Unit, University Hospital of Siena, Siena, Italy
  • Francesca Montagnani; Department of Medical Biotechnologies, University of Siena, Siena, Italy; Department of Specialized and Internal Medicine, Tropical and Infectious Diseases Unit
  • Emanuele Montomoli; VisMederi S.r.l, Siena, Italy; VisMederi Research S.r.l., Siena, Italy; Department of Molecular and Developmental Medicine, University of Siena, Siena, Italy
  • Claudia Sala; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy
  • Giuseppe Ippolito; National Institute for Infectious Diseases Lazzaro Spallanzani, IRCCS, Rome, Italy
  • Rino Rappuoli; Monoclonal Antibody Discovery (MAD) Lab, Fondazione Toscana Life Sciences, Siena, Italy; Faculty of Medicine, Imperial College, London, United Kingdom
Preprint in English | bioRxiv | ID: ppbiorxiv-078154
ABSTRACT
In the absence of approved drugs or vaccines, there is a pressing need to develop tools for therapy and prevention of Covid-19. Human monoclonal antibodies have very good probability of being safe and effective tools for therapy and prevention of SARS-CoV-2 infection and disease. Here we describe the screening of PBMCs from seven people who survived Covid-19 infection to isolate human monoclonal antibodies against SARS-CoV-2. Over 1,100 memory B cells were single-cell sorted using the stabilized prefusion form of the spike protein and incubated for two weeks to allow natural production of antibodies. Supernatants from each cell were tested by ELISA for spike protein binding, and positive antibodies were further tested for neutralization of spike binding to receptor(s) on Vero E6 cells and for virus neutralization in vitro. From the 1,167 memory B specific for SARS-CoV-2, we recovered 318 B lymphocytes expressing human monoclonals recognizing the spike protein and 74 of these were able to inhibit the binding of the spike protein to the receptor. Finally, 17 mAbs were able to neutralize the virus when assessed for neutralization in vitro. Lead candidates to progress into the drug development pipeline will be selected from the panel of neutralizing antibodies identified with the procedure described in this study. One Sentence SummaryNeutralizing human monoclonal antibodies isolated from Covid-19 convalescent patients for therapeutic and prophylactic interventions.
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Full text: Available Collection: Preprints Database: bioRxiv Type of study: Prognostic study Language: English Year: 2020 Document type: Preprint
Full text: Available Collection: Preprints Database: bioRxiv Type of study: Prognostic study Language: English Year: 2020 Document type: Preprint
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