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© Christophe Thomas, Institut Pasteur
Gel d’électrophorèse lors de la purification d’un échantillon protéique
Publication : The Journal of biological chemistry

Development of a highly specific and sensitive VHH-based sandwich immunoassay for the detection of the SARS-CoV-2 nucleoprotein.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in The Journal of biological chemistry - 19 Oct 2021

Gransagne M, Aymé G, Brier S, Chauveau-Le Friec G, Meriaux V, Nowakowski M, Dejardin F, Levallois S, Dias de Melo G, Donati F, Prot M, Brûlé S, Raynal B, Bellalou J, Goncalves P, Montagutelli X, Di Santo JP, Lazarini F, England P, Petres S, Escriou N, Lafaye P,

Link to Pubmed [PMID] – 34678315

Link to DOI – S0021-9258(21)01093-010.1016/j.jbc.2021.101290

J Biol Chem 2021 Oct; (): 101290

The current COVID-19 pandemic illustrates the importance of obtaining reliable methods for the rapid detection of SARS-CoV-2. A highly specific and sensitive diagnostic test able to differentiate the SARS-CoV-2 virus from common human coronaviruses is therefore needed. Coronavirus nucleoprotein (N) localizes to the cytoplasm and the nucleolus, and are required for viral RNA synthesis. N is the most abundant coronavirus protein, so it is of utmost importance to develop specific antibodies for its detection. In this study, we developed a sandwich immunoassay to recognize the SARS-CoV-2 N protein. We immunized one alpaca with recombinant SARS-CoV-2 N and constructed a large single variable domain on heavy chain (VHH) antibody library. After phage display selection, 7 VHHs recognizing the full N protein were identified by ELISA. These VHHs did not recognize the nucleoproteins of the four common human coronaviruses. Hydrogen Deuterium eXchange-Mass Spectrometry (HDX-MS) analysis also showed that these VHHs mainly targeted conformational epitopes in either the C-terminal or the N-terminal domains. All VHHs were able to recognize SARS-CoV-2 in infected cells or on infected hamster tissues. Moreover, the VHHs could detect the SARS variants B.1.17/alpha, B.1.351/beta and P1/gamma. We propose that this sandwich immunoassay could be applied to specifically detect the SARS-CoV-2 N in human nasal swabs.

https://pubmed.ncbi.nlm.nih.gov/34678315