SARS-CoV-2 Spike protein peptides displayed in the Pyrococcus furiosus RAD system preserve epitopes antigenicity, immunogenicity, and virus-neutralizing activity of antibodies

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Cioffi, Victor Bolsanelli  ORCID logo
de Castro-Amarante, Maria Fernanda  ORCID logo
Andreata-Santos, Robert  ORCID logo

Amongst the potential contribution of protein or peptide-display systems to study epitopes with relevant immunological features, the RAD display system stands out as a highly stable scaffold protein that allows the presentation of constrained target peptides. Here, we employed the RAD display system to present peptides derived from the SARS-CoV-2 Spike (S) protein as a tool to detect specific serum antibodies and to generate polyclonal antibodies capable of inhibiting SARS-CoV-2 infectivity in vitro. 44 linear S-derived peptides were genetically fused with the RAD scaffold (RAD-SCoV-epitopes) and screened for antigenicity with sera collected from COVID-19-infected patients. In a second step, selected RAD-SCoV-epitopes were used to immunize mice and generate antibodies. Phenotypic screening showed that some of these antibodies were able to recognize replicating viral particles in VERO CCL-81 and most notably seven of the RAD-SCoV-epitopes were able to induce antibodies that inhibited viral infection. Our findings highlight the RAD display system as an useful platform for the immunological characterization of peptides and a potentially valuable strategy for the design of antigens for peptide-based vaccines, for epitope-specific antibody mapping, and for the development of antibodies for diagnostic and therapeutic purposes.


Acknowledgements: We thank the Department of Microbiology (USP) for support and access to instrumentation and the Centre of Facility for Research from the Institute of Biomedical Sciences (CEFAP) from University of São Paulo.

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Scientific Reports
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Nature Publishing Group UK
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (88881.507138/2020-01, 88887.505029/2020-00, 88887.505029/2020-00, 88887.505029/2020-00, 88887.505029/2020-00, 88881.507138/2020-01, 88887.505029/2020-00, 88881.507138/2020-01)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (4015/0620207, 4015/0620207, 308650/2019-0, 308650/2019-0, 308650/2019-0, 308650/2019-0)
Global Challenge Research Funding (RG102642, RG102642, RG102642, RG102642, RG102642, RG102642)