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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2021
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.
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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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