Development of an anti‐SARS‐CoV‐2 monoclonal antibody panel and its applicability as a reagent in high‐throughput fluorescence reduction neutralization and immunohistochemistry assays

Development of an anti‐SARS‐CoV‐2 monoclonal antibody panel and its applicability as a reagent in high‐throughput fluorescence reduction neutralization and immunohistochemistry assays

Abstract Since its emergence in late 2019, coronavirus disease 2019 (COVID‐19) has caused millions of deaths and socioeconomic losses. Although vaccination significantly reduced disease mortality, it has been shown that protection wanes over time, and that severe acute respiratory syndrome coronavir...

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Published in:Journal of medical virology Vol. 95; no. 9; p. e29111
Main Authors: Coelho, Gabriela M., Cataneo, Allan H. D., Raboni, Sonia M., Nogueira, Meri B., de Paula, Caroline B. Vaz, Almeida, Ana C. S. F., Rogerio, Vanessa Z., Zanchin, Nilson T., de Noronha, Lucia, Zanluca, Camila, Duarte dos Santos, Claudia N.
Format: Journal Article
Language:English
Published: London Wiley Subscription Services, Inc 01-09-2023
Subjects:
Assaying
Coronaviruses
COVID-19
COVID-19 vaccines
Fluorescence
Immunity
Immunohistochemistry
Monoclonal antibodies
Neutralization
Pandemics
Reagents
Reduction
Respiratory diseases
Severe acute respiratory syndrome
Severe acute respiratory syndrome coronavirus 2
Vaccines
Viral diseases
Virology
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Summary:Abstract Since its emergence in late 2019, coronavirus disease 2019 (COVID‐19) has caused millions of deaths and socioeconomic losses. Although vaccination significantly reduced disease mortality, it has been shown that protection wanes over time, and that severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) variants of concern (VOCs) may escape vaccine‐derived immunity. Therefore, serological studies are necessary to assess protection in the population and guide vaccine regimens. A common measure of protective immunity is the presence of neutralizing antibodies (nAbs). However, the gold standard for measuring nAbs (plaque reduction neutralization test, or PRNT) is laborious and time‐consuming, limiting its large‐scale applicability. We developed a high‐throughput fluorescence reduction neutralization assay (FRNA) to detect SARS‐CoV‐2 nAbs. Because the assay relies on immunostaining, we developed and characterized monoclonal antibodies (mAbs) to lower costs and reduce the assay's vulnerability to reagent shortages. Using samples of individuals vaccinated with COVID‐19 and unvaccinated/pre‐pandemic samples, we showed that FRNA results using commercial and in‐house mAbs strongly correlated with those of the PRNT method while providing results in 70% less time. In addition to providing a fast, reliable, and high‐throughput alternative for measuring nAbs, the FRNA can be easily customized to assess SARS‐CoV‐2 VOCs. Additionally, the mAb we produced was able to detect SARS‐CoV‐2 in pulmonary tissues by immunohistochemistry assays.
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ISSN:0146-6615
1096-9071
DOI:10.1002/jmv.29111