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Front Immunol . A novel precision-serology assay for SARS-CoV-2 infection based on linear B-cell epitopes of Spike protein

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  • Front Immunol . A novel precision-serology assay for SARS-CoV-2 infection based on linear B-cell epitopes of Spike protein

    Front Immunol


    . 2023 May 12;14:1166924.
    doi: 10.3389/fimmu.2023.1166924. eCollection 2023. A novel precision-serology assay for SARS-CoV-2 infection based on linear B-cell epitopes of Spike protein

    Samuel B Lundin 1 2 3 , Hanna Kann 1 , Alma Fulurija 2 3 4 , Björn Andersson 1 , Sravya S Nakka 1 , Lars-Magnus Andersson 5 6 , Magnus Gisslén 5 6 , Ali M Harandi 1 7



    AffiliationsAbstract

    Introduction: The COVID-19 pandemic illustrates the need for serology diagnostics with improved accuracy. While conventional serology based on recognition of entire proteins or subunits thereof has made significant contribution to the antibody assessment space, it often suffers from sub-optimal specificity. Epitope-based, high-precision, serology assays hold potential to capture the high specificity and diversity of the immune system, hence circumventing the cross-reactivity with closely related microbial antigens.
    Methods: We herein report mapping of linear IgG and IgA antibody epitopes of the SARS-CoV-2 Spike (S) protein in samples from SARS-CoV-2 exposed individuals along with certified SARS-CoV-2 verification plasma samples using peptide arrays.
    Results: We identified 21 distinct linear epitopes. Importantly, we showed that pre-pandemic serum samples contain IgG antibodies reacting to the majority of protein S epitopes, most likely as a result of prior infection with seasonal coronaviruses. Only 4 of the identified SARS-CoV-2 protein S linear epitopes were specific for SARS-CoV-2 infection. These epitopes are located at positions 278-298 and 550-586, just proximal and distal to the RBD, as well as at position 1134-1156 in the HR2 subdomain and at 1248-1271 in the C-terminal subdomain of protein S. To substantiate the applicability of our findings, we tested three of the high-accuracy protein S epitopes in a Luminex assay, using a certified validation plasma sample set from SARS-CoV-2 infected individuals. The Luminex results were well aligned with the peptide array results, and correlated very well with in-house and commercial immune assays for RBD, S1 and S1/S2 domains of protein S.
    Conclusion: We present a comprehensive mapping of linear B-cell epitopes of SARS-CoV-2 protein S, that identifies peptides suitable for a precision serology assay devoid of cross-reactivity. These results have implications for development of highly specific serology test for exposure to SARS-CoV-2 and other members of the coronaviridae family, as well as for rapid development of serology tests for future emerging pandemic threats.

    Keywords: B-cell epitope; SARS-CoV-2; Spike protein; cross-reactivity; precision serology.

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