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Article

SARS-CoV-2 Epitope Mapping on Microarrays Highlights Strong Immune-Response to N Protein Region

1
National Research Council of Italy, Istituto di Scienze e Tecnologie Chimiche “Giulio Natta” (SCITEC-CNR), Via Mario Bianco 9, 20131 Milano, Italy
2
Department of Molecular Medicine, University of Padova, Via A. Gabelli 63, 35121 Padova, Italy
3
Department of Infectious-Tropical Diseases and Microbiology, IRCCS Sacro Cuore Don Calabria Hospital, Via Don A. Sempreboni, 5-37024 Negrar di Valpolicella, Italy
*
Authors to whom correspondence should be addressed.
These authors equally contributed to the paper.
These authors equally contributed to the paper.
Vaccines 2021, 9(1), 35; https://doi.org/10.3390/vaccines9010035
Received: 11 December 2020 / Revised: 6 January 2021 / Accepted: 6 January 2021 / Published: 11 January 2021
A workflow for rapid SARS-CoV-2 epitope discovery on peptide microarrays is herein reported. The process started with a proteome-wide screening of immunoreactivity based on the use of a high-density microarray followed by a refinement and validation phase on a restricted panel of probes using microarrays with tailored peptide immobilization through a click-based strategy. Progressively larger, independent cohorts of Covid-19 positive sera were tested in the refinement processes, leading to the identification of immunodominant regions on SARS-CoV-2 spike (S), nucleocapsid (N) protein and Orf1ab polyprotein. A summary study testing 50 serum samples highlighted an epitope of the N protein (region 155–71) providing good diagnostic performance in discriminating Covid-19 positive vs. healthy individuals. Using this epitope, 92% sensitivity and 100% specificity were reached for IgG detection in Covid-19 samples, and no cross-reactivity with common cold coronaviruses was detected. Likewise, IgM immunoreactivity in samples collected within the first month after symptoms onset showed discrimination ability. Overall, epitope 155–171 from N protein represents a promising candidate for further development and rapid implementation in serological tests. View Full-Text
Keywords: SARS-CoV-2; Covid-19; serological test; epitope mapping; peptide microarrays; click chemistry SARS-CoV-2; Covid-19; serological test; epitope mapping; peptide microarrays; click chemistry
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MDPI and ACS Style

Musicò, A.; Frigerio, R.; Mussida, A.; Barzon, L.; Sinigaglia, A.; Riccetti, S.; Gobbi, F.; Piubelli, C.; Bergamaschi, G.; Chiari, M.; Gori, A.; Cretich, M. SARS-CoV-2 Epitope Mapping on Microarrays Highlights Strong Immune-Response to N Protein Region. Vaccines 2021, 9, 35. https://doi.org/10.3390/vaccines9010035

AMA Style

Musicò A, Frigerio R, Mussida A, Barzon L, Sinigaglia A, Riccetti S, Gobbi F, Piubelli C, Bergamaschi G, Chiari M, Gori A, Cretich M. SARS-CoV-2 Epitope Mapping on Microarrays Highlights Strong Immune-Response to N Protein Region. Vaccines. 2021; 9(1):35. https://doi.org/10.3390/vaccines9010035

Chicago/Turabian Style

Musicò, Angelo, Roberto Frigerio, Alessandro Mussida, Luisa Barzon, Alessandro Sinigaglia, Silvia Riccetti, Federico Gobbi, Chiara Piubelli, Greta Bergamaschi, Marcella Chiari, Alessandro Gori, and Marina Cretich. 2021. "SARS-CoV-2 Epitope Mapping on Microarrays Highlights Strong Immune-Response to N Protein Region" Vaccines 9, no. 1: 35. https://doi.org/10.3390/vaccines9010035

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