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Abstract
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.
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1 University of São Paulo, Laboratory of Applied Structural Biology, Department of Microbiology, Institute of Biomedical Sciences, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722)
2 University of São Paulo, Institute of Biomedical Sciences, Laboratory of Vaccine Development, Department of Microbiology, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722)
3 University of Cambridge, Department of Biochemistry, Cambridge, UK (GRID:grid.5335.0) (ISNI:0000 0001 2188 5934)
4 University of São Paulo, Av. Prof. Lineu Prestes, Core Facilities to Support Research (CEFAP), Institute of Biomedical Sciences, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722)
5 University of São Paulo, Institute of Biomedical Sciences, Laboratory of Vaccine Development, Department of Microbiology, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722); Butantan Institute, Vaccine Development Laboratory, São Paulo, Brazil (GRID:grid.418514.d) (ISNI:0000 0001 1702 8585)
6 University of São Paulo, Institute of Biomedical Sciences, Phenotypic Screening Platform, Department of Microbiology, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722)
7 University of São Paulo, Laboratory of Clinical and Molecular Virology, Institute of Biomedical Sciences, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722); Institut Pasteur de São Paulo, São Paulo, Brazil (GRID:grid.11899.38)
8 University of São Paulo, Institute of Biomedical Sciences, Laboratory of Vaccine Development, Department of Microbiology, São Paulo, Brazil (GRID:grid.11899.38) (ISNI:0000 0004 1937 0722); Institut Pasteur de São Paulo, São Paulo, Brazil (GRID:grid.11899.38)