Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples

Mazzini, L.; Martinuzzi, D.; Hyseni, I.; Lapini, G.; Benincasa, L.; Piu, P.; Trombetta, C. M.; Marchi, S.; Razzano, I.; Manenti, A.; Montomoli, E.

doi:10.1101/2020.08.10.243717

bioRxiv · 10.1101/2020.08.10.243717

Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples

Mazzini, L.·Martinuzzi, D.·Hyseni, I.·Lapini, G.·Benincasa, L.·Piu, P.·Trombetta, C. M.·Marchi, S.·Razzano, I.·Manenti, A.·Montomoli, E.

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Abstract

A newly identified coronavirus, named SARS-CoV-2, emerged in December 2019 in Hubei Province, China, and quickly spread throughout the world; so far, it has caused more than 18 million cases of disease and 700,000 deaths. The diagnosis of SARS-CoV-2 infection is currently based on the detection of viral RNA in nasopharyngeal swabs by means of molecular-based assays, such as real-time RT-PCR. Furthermore, serological assays aimed at detecting different classes of antibodies constitute the best surveillance strategy for gathering information on the humoral immune response to infection and the spread of the virus through the population, in order to evaluate the immunogenicity of novel future vaccines and medicines for the treatment and prevention of COVID-19 disease. The aim of this study was to determine SARS-CoV-2-specific antibodies in human serum samples by means of different commercial and in-house ELISA kits, in order to evaluate and compare their results first with one another and then with those yielded by functional assays using wild-type virus. It is important to know the level of SARS-CoV-2-specific IgM, IgG and IgA antibodies in order to predict population immunity and possible cross-reactivity with other coronaviruses and to identify potentially infectious subjects. In addition, in a small sub-group of samples, we performed a subtyping Immunoglobulin G ELISA. Our data showed an excellent statistical correlation between the neutralization titer and the IgG, IgM and IgA ELISA response against the receptor-binding domain of the spike protein, confirming that antibodies against this portion of the virus spike protein are highly neutralizing and that the ELISA Receptor-Binding Domain-based assay can be used as a valid surrogate for the neutralization assay in laboratories which do not have Biosecurity level-3 facilities.

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Mazzini, L., Martinuzzi, D., Hyseni, I., Lapini, G., Benincasa, L., Piu, P., Trombetta, C. M., Marchi, S., Razzano, I., Manenti, A., Montomoli, E.. 2020-08-10. Comparative analyses of SARS-CoV-2 binding (IgG, IgM, IgA) and neutralizing antibodies from human serum samples. https://doi.org/10.1101/2020.08.10.243717

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