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Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture

Determining the presence of antibodies to the SARS-CoV-2 antigens is the best way to identify infected people, regardless of the development of symptoms of COVID-19. The nucleoprotein (NP) of the SARS-CoV-2 is an immunodominant antigen of the virus; anti-NP antibodies are detected in persons previou...

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Autores principales: Kolesov, Denis E., Sinegubova, Maria V., Safenkova, Irina V., Vorobiev, Ivan I., Orlova, Nadezhda A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8744485/
https://www.ncbi.nlm.nih.gov/pubmed/35036106
http://dx.doi.org/10.7717/peerj.12751
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author Kolesov, Denis E.
Sinegubova, Maria V.
Safenkova, Irina V.
Vorobiev, Ivan I.
Orlova, Nadezhda A.
author_facet Kolesov, Denis E.
Sinegubova, Maria V.
Safenkova, Irina V.
Vorobiev, Ivan I.
Orlova, Nadezhda A.
author_sort Kolesov, Denis E.
collection PubMed
description Determining the presence of antibodies to the SARS-CoV-2 antigens is the best way to identify infected people, regardless of the development of symptoms of COVID-19. The nucleoprotein (NP) of the SARS-CoV-2 is an immunodominant antigen of the virus; anti-NP antibodies are detected in persons previously infected with the virus with the highest titers. Many test systems for detecting antibodies to SARS-CoV-2 contain NP or its fragments as antigen. The sensitivity and specificity of such test systems differ significantly, which can be explained by variations in the antigenic properties of NP caused by differences in the methods of its cultivation, isolation and purification. We investigated this effect for the Escherichia coli-derived SARS-CoV-2 NP, obtained from the cytoplasm in the soluble form. We hypothesized that co-purified nucleic acids that form a strong complex with NP might negatively affect NP’s antigenic properties. Therefore, we have established the NP purification method, which completely eliminates the RNA in the NP preparation. Two stages of RNA removal were used: treatment of the crude lysate of E. coli with RNase A and subsequent selective RNA elution with 2 M NaCl solution. The resulting NP without RNA has a significantly better signal-to-noise ratio when used as an ELISA antigen and tested with a control panel of serum samples with antibodies to SARS-CoV-2; therefore, it is preferable for in vitro diagnostic use. The same increase of the signal-to-noise ratio was detected for the free N-terminal domain of the NP. Complete removal of RNA complexed with NP during purification will significantly improve its antigenic properties, and the absence of RNA in NP preparations should be controlled during the production of this antigen.
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spelling pubmed-87444852022-01-14 Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture Kolesov, Denis E. Sinegubova, Maria V. Safenkova, Irina V. Vorobiev, Ivan I. Orlova, Nadezhda A. PeerJ Biotechnology Determining the presence of antibodies to the SARS-CoV-2 antigens is the best way to identify infected people, regardless of the development of symptoms of COVID-19. The nucleoprotein (NP) of the SARS-CoV-2 is an immunodominant antigen of the virus; anti-NP antibodies are detected in persons previously infected with the virus with the highest titers. Many test systems for detecting antibodies to SARS-CoV-2 contain NP or its fragments as antigen. The sensitivity and specificity of such test systems differ significantly, which can be explained by variations in the antigenic properties of NP caused by differences in the methods of its cultivation, isolation and purification. We investigated this effect for the Escherichia coli-derived SARS-CoV-2 NP, obtained from the cytoplasm in the soluble form. We hypothesized that co-purified nucleic acids that form a strong complex with NP might negatively affect NP’s antigenic properties. Therefore, we have established the NP purification method, which completely eliminates the RNA in the NP preparation. Two stages of RNA removal were used: treatment of the crude lysate of E. coli with RNase A and subsequent selective RNA elution with 2 M NaCl solution. The resulting NP without RNA has a significantly better signal-to-noise ratio when used as an ELISA antigen and tested with a control panel of serum samples with antibodies to SARS-CoV-2; therefore, it is preferable for in vitro diagnostic use. The same increase of the signal-to-noise ratio was detected for the free N-terminal domain of the NP. Complete removal of RNA complexed with NP during purification will significantly improve its antigenic properties, and the absence of RNA in NP preparations should be controlled during the production of this antigen. PeerJ Inc. 2022-01-07 /pmc/articles/PMC8744485/ /pubmed/35036106 http://dx.doi.org/10.7717/peerj.12751 Text en ©2022 Kolesov et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Biotechnology
Kolesov, Denis E.
Sinegubova, Maria V.
Safenkova, Irina V.
Vorobiev, Ivan I.
Orlova, Nadezhda A.
Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture
title Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture
title_full Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture
title_fullStr Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture
title_full_unstemmed Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture
title_short Antigenic properties of the SARS-CoV-2 nucleoprotein are altered by the RNA admixture
title_sort antigenic properties of the sars-cov-2 nucleoprotein are altered by the rna admixture
topic Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8744485/
https://www.ncbi.nlm.nih.gov/pubmed/35036106
http://dx.doi.org/10.7717/peerj.12751
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