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Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies
The development of new diagnostic assays become a priority for managing COVID-19. To this aim, we presented here an in-house ELISA based on the production of two major recombinant and high-quality antigens from SARS-CoV-2. Full-length N and S-RBD fragment proteins fused to mouse IgG2a-Fc were produc...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152238/ https://www.ncbi.nlm.nih.gov/pubmed/34051226 http://dx.doi.org/10.1016/j.jim.2021.113082 |
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author | Frumence, Etienne Lebeau, Grégorie Viranaicken, Wildriss Dobi, Anthony Vagner, Damien Lalarizo Rakoto, Mahary Sandenon Seteyen, Anne-Laure Giry, Claude Septembre-Malaterre, Axelle Raffray, Loïc Gasque, Philippe |
author_facet | Frumence, Etienne Lebeau, Grégorie Viranaicken, Wildriss Dobi, Anthony Vagner, Damien Lalarizo Rakoto, Mahary Sandenon Seteyen, Anne-Laure Giry, Claude Septembre-Malaterre, Axelle Raffray, Loïc Gasque, Philippe |
author_sort | Frumence, Etienne |
collection | PubMed |
description | The development of new diagnostic assays become a priority for managing COVID-19. To this aim, we presented here an in-house ELISA based on the production of two major recombinant and high-quality antigens from SARS-CoV-2. Full-length N and S-RBD fragment proteins fused to mouse IgG2a-Fc were produced in the CHO cell line. Secreted recombinant proteins were easily purified with standard Protein A chromatography and were used in an in-house ELISA to detect anti-N and anti-RBD IgGs in the plasma of COVID-19 RTPCR-positive patients. High reactivity against recombinant antigens was readily detected in all positive plasma samples, whereas no recognition was observed with control healthy subject's plasmas. Remarkably, unpurified recombinant N protein obtained from cell culture supernatant was also suitable for the monitoring by ELISA of IgG levels in positive patients. This work provides an early prospection for low price but high-quality serological kit development. |
format | Online Article Text |
id | pubmed-8152238 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-81522382021-05-28 Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies Frumence, Etienne Lebeau, Grégorie Viranaicken, Wildriss Dobi, Anthony Vagner, Damien Lalarizo Rakoto, Mahary Sandenon Seteyen, Anne-Laure Giry, Claude Septembre-Malaterre, Axelle Raffray, Loïc Gasque, Philippe J Immunol Methods Technical Note The development of new diagnostic assays become a priority for managing COVID-19. To this aim, we presented here an in-house ELISA based on the production of two major recombinant and high-quality antigens from SARS-CoV-2. Full-length N and S-RBD fragment proteins fused to mouse IgG2a-Fc were produced in the CHO cell line. Secreted recombinant proteins were easily purified with standard Protein A chromatography and were used in an in-house ELISA to detect anti-N and anti-RBD IgGs in the plasma of COVID-19 RTPCR-positive patients. High reactivity against recombinant antigens was readily detected in all positive plasma samples, whereas no recognition was observed with control healthy subject's plasmas. Remarkably, unpurified recombinant N protein obtained from cell culture supernatant was also suitable for the monitoring by ELISA of IgG levels in positive patients. This work provides an early prospection for low price but high-quality serological kit development. Elsevier B.V. 2021-08 2021-05-26 /pmc/articles/PMC8152238/ /pubmed/34051226 http://dx.doi.org/10.1016/j.jim.2021.113082 Text en © 2021 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Technical Note Frumence, Etienne Lebeau, Grégorie Viranaicken, Wildriss Dobi, Anthony Vagner, Damien Lalarizo Rakoto, Mahary Sandenon Seteyen, Anne-Laure Giry, Claude Septembre-Malaterre, Axelle Raffray, Loïc Gasque, Philippe Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies |
title | Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies |
title_full | Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies |
title_fullStr | Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies |
title_full_unstemmed | Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies |
title_short | Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies |
title_sort | robust and low-cost elisa based on igg-fc tagged recombinant proteins to screen for anti-sars-cov-2 antibodies |
topic | Technical Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152238/ https://www.ncbi.nlm.nih.gov/pubmed/34051226 http://dx.doi.org/10.1016/j.jim.2021.113082 |
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