tetano
Editor, Senior Moderator
Methods Mol Biol
. 2023;2578:209-217.
doi: 10.1007/978-1-0716-2732-7_15.
Epitope Mapping on Microarrays Highlights a Sequence on the N Protein with Strong Immune Response in SARS-CoV-2 Patients
Roberto Frigerio[SUP] 1 2 [/SUP], Angelo Musicò[SUP] 3 4 [/SUP], Alessandro Strada[SUP] 3 5 [/SUP], Alessandro Mussida[SUP] 3 6 [/SUP], Paola Gagni[SUP] 3 [/SUP], Greta Bergamaschi[SUP] 3 [/SUP], Marcella Chiari[SUP] 3 [/SUP], Luisa Barzon[SUP] 7 [/SUP], Alessandro Gori[SUP] 3 [/SUP], Marina Cretich[SUP] 3 [/SUP]
Affiliations
Abstract
In SARS-CoV-2 pandemic scenario, the identification of rapid methods to detect antibodies against coronavirus has been a wide and urgent issue. Epitope mapping on peptide microarrays is a rapid way to identify sequences with a high immunoreactivity. The process begins with a proteome-wide screening, based on immune affinity; the use of a high-density microarray is followed by a validation phase, where a restricted panel of probes is tested using peptide microarrays; peptide sequences are immobilized through a click-based strategy.COVID-19-positive sera are tested and immuno-domains regions are identified on SARS-CoV-2 spike (S), nucleocapsid (N) protein, and Orf1ab polyprotein. An epitope on N protein (region 155-171) provided good diagnostic performance in discriminating COVID-19-positive vs. healthy individuals. Using this sequence, 92% sensitivity and 100% specificity are reached for IgG detection in COVID-19 samples, and no cross-reactivity with common cold coronaviruses is detected. Overall, epitope 155-171 from N protein represents a promising candidate for further development and rapid implementation in serological tests.
Keywords: Epitope mapping; Immune-response; Peptide microarrays; SARS-CoV-2.
. 2023;2578:209-217.
doi: 10.1007/978-1-0716-2732-7_15.
Epitope Mapping on Microarrays Highlights a Sequence on the N Protein with Strong Immune Response in SARS-CoV-2 Patients
Roberto Frigerio[SUP] 1 2 [/SUP], Angelo Musicò[SUP] 3 4 [/SUP], Alessandro Strada[SUP] 3 5 [/SUP], Alessandro Mussida[SUP] 3 6 [/SUP], Paola Gagni[SUP] 3 [/SUP], Greta Bergamaschi[SUP] 3 [/SUP], Marcella Chiari[SUP] 3 [/SUP], Luisa Barzon[SUP] 7 [/SUP], Alessandro Gori[SUP] 3 [/SUP], Marina Cretich[SUP] 3 [/SUP]
Affiliations
- PMID: 36152290
- DOI: 10.1007/978-1-0716-2732-7_15
Abstract
In SARS-CoV-2 pandemic scenario, the identification of rapid methods to detect antibodies against coronavirus has been a wide and urgent issue. Epitope mapping on peptide microarrays is a rapid way to identify sequences with a high immunoreactivity. The process begins with a proteome-wide screening, based on immune affinity; the use of a high-density microarray is followed by a validation phase, where a restricted panel of probes is tested using peptide microarrays; peptide sequences are immobilized through a click-based strategy.COVID-19-positive sera are tested and immuno-domains regions are identified on SARS-CoV-2 spike (S), nucleocapsid (N) protein, and Orf1ab polyprotein. An epitope on N protein (region 155-171) provided good diagnostic performance in discriminating COVID-19-positive vs. healthy individuals. Using this sequence, 92% sensitivity and 100% specificity are reached for IgG detection in COVID-19 samples, and no cross-reactivity with common cold coronaviruses is detected. Overall, epitope 155-171 from N protein represents a promising candidate for further development and rapid implementation in serological tests.
Keywords: Epitope mapping; Immune-response; Peptide microarrays; SARS-CoV-2.