Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies

Frumence, Étienne; Lebeau, Grégorie; Viranaïcken, Wildriss; Dobi, Anthony; Vagner, Damien; Rakoto, Mahary Lalarizo; Seteyen, Anne-Laure Sandenon; Giry, Claude; Septembre-Malaterre, Axelle; Raffray, L.; Gasque, Philippe

doi:10.1016/j.jim.2021.113082

Cited by 6 publications

(4 citation statements)

References 15 publications

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“…At the international level, similar studies describing COVID-19 immunoassays have been reported from several research groups [ 29 , 30 , 31 , 32 , 33 , 34 ]. Most of these immunoassays were based on SARS-CoV-2 full-length S, NP, or fragments of S (e.g., receptor binding domain).…”

Section: Development Of Serological Assays For Covid-19supporting

confidence: 54%

“…In some cases, assays were optimized not only for detection of IgM and IgG, but also for IgA [ 33 ]. Validation of results was also evaluated against RT-PCR, the MN assay, and commercially available kits [ 29 , 30 , 31 , 32 , 33 , 34 ]. Herein, we review several in-house immunoassays developed in Saudi Arabia.…”

Section: Development Of Serological Assays For Covid-19mentioning

confidence: 99%

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Development of Serological Assays and Seroprevalence Studies of the New Coronavirus 2019 (COVID-19): Reports from Saudi Arabia

Alandijany

Faizo

2021

Healthcare

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Serological assays are valuable tools for tracking COVID-19 spread, estimation of herd immunity, and evaluation of vaccine effectiveness. Several reports from Saudi Arabia describe optimized in-house protocols that enable detection of SARS-CoV-2 specific antibodies and measurement of their neutralizing activity. Notably, there were variations in the approaches utilized to develop and validate these immunoassays in term of sample size, validation methodologies, and statistical analyses. The developed enzyme-linked immunoassays (ELISAs) were based on the viral full-length spike (S), S1 subunit, and nucleocapsid (NP), and enabled detection of IgM and/or IgG. ELISAs were evaluated and validated against a microneutralization assay utilizing a local SARS-CoV-2 clinical isolate, FDA-approved commercially available immunoassays, and/or real-time polymerase chain reaction (RT-PCR). Overall, the performance of the described assays was high, reaching up to 100% sensitivity and 98.9% specificity with no cross-reactivity with other coronaviruses. In-house immunoassays, along with commercially available kits, were subsequently applied in a number of sero-epidemiological studies aiming to estimate sero-positivity status among local populations including healthcare workers, COVID-19 patients, non-COVID-19 patients, and healthy blood donors. The reported seroprevalence rates differed widely among these studies, ranging from 0.00% to 32.2%. These variations are probably due to study period, targeted population, sample size, and performance of the immunoassays utilized. Indeed, lack of sero-positive cases were reported among healthy blood donors during the lockdown, while the highest rates were reported when the number of COVID-19 cases peaked in the country, particularly among healthcare workers working in referral hospitals and quarantine sites. In this review, we aim to (1) provide a critical discussion about the developed in-house immunoassays, and (2) summarize key findings of the sero-epidemiological studies and highlight strengths and weaknesses of each study.

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Section: Development Of Serological Assays For Covid-19supporting

confidence: 54%

Section: Development Of Serological Assays For Covid-19mentioning

confidence: 99%

Development of Serological Assays and Seroprevalence Studies of the New Coronavirus 2019 (COVID-19): Reports from Saudi Arabia

Alandijany

Faizo

2021

Healthcare

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“…The RBD, as an isolated protein, is broadly used in different types of clinical and medical applications (serological tests, vaccine formulation, etc.) [ 17 , 18 , 19 , 20 ]; therefore, its in vitro production is of paramount importance. Mammalian and insect cells are the model systems of election used for the heterologous expression of SARS-CoV-2 RBD due to its intrinsic structural complexity.…”

Section: Introductionmentioning

confidence: 99%

The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression

Maffei¹,

Montemiglio

Vitagliano³

et al. 2021

Biomolecules

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COVID-19 is a highly infectious disease caused by a newly emerged coronavirus (SARS-CoV-2) that has rapidly progressed into a pandemic. This unprecedent emergency has stressed the significance of developing effective therapeutics to fight the current and future outbreaks. The receptor-binding domain (RBD) of the SARS-CoV-2 surface Spike protein is the main target for vaccines and represents a helpful “tool” to produce neutralizing antibodies or diagnostic kits. In this work, we provide a detailed characterization of the native RBD produced in three major model systems: Escherichia coli, insect and HEK-293 cells. Circular dichroism, gel filtration chromatography and thermal denaturation experiments indicated that recombinant SARS-CoV-2 RBD proteins are stable and correctly folded. In addition, their functionality and receptor-binding ability were further evaluated through ELISA, flow cytometry assays and bio-layer interferometry.

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“…Mammalian codon-optimized genes coding either for the SARS-CoV-2 N protein or the RBD domain of spike protein (residues 328/583) (UniProtKB-P0DTC9 (NCAP_SARS2)) were established using Cricetulus griseus codon usage as reference 40 . The sequence encoding the mouse Ig gamma-2A (IgG2A) chain C region sequence (UniprotKB-P01863 (GCAA_MOUSE) was fused in frame to the carboxy-terminus of N and RBD proteins with a spacer serine-glycine spacer.…”

Section: Methodsmentioning

confidence: 99%

The SARS-CoV-2 spike residues 616/644 and 1138/1169 delineate two antibody epitopes in COVID-19 mRNA COMIRNATY vaccine (Pfizer/BioNTech)

Andries

Viranaïcken

Cordonin³

et al. 2022

Sci Rep

Self Cite

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The newly identified coronavirus SARS-CoV-2 is responsible for the worldwide pandemic COVID-19. Considerable efforts have been devoted for the development of effective vaccine strategies against COVID-19. The SARS-CoV-2 spike protein has been identified as the major antigen candidate for the development of COVID-19 vaccines. The Pfizer-BioNTech COVID-19 vaccine comirnaty is a lipid nanoparticle-encapsulated mRNA encoding a full-length and prefusion-stabilized SARS-CoV-2 spike protein. In the present study, synthetic peptide-based ELISA assays were performed to identify linear B-cell epitopes into the spike protein that contribute to elicitation of antibody response in comirnaty-vaccinated individuals. The synthetic S2P6 peptide containing the spike residues 1138/1169 and to a lesser extent, the synthetic S1P4 peptide containing the spike residues 616/644 were recognized by the immune sera from comirnaty vaccine recipients but not COVID-19 recovered patients. We assume that the synthetic S2P6 peptide and to a lesser extent the synthetic S1P4 peptide, could be of interest to measure the dynamic of antibody response to COVID-19 mRNA vaccines. The S2P6 peptide has been identified as immunogenic in adult BALB/c mice that received protein-peptide conjugates in a prime-boost schedule. This raises the question on the role of the B-cell epitope peptide containing the SARS-CoV-2 spike residues 1138/1169 in protective efficacy of the Pfizer-BioNTech COVID-19 vaccine comirnaty.

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Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies

Cited by 6 publications

References 15 publications

Development of Serological Assays and Seroprevalence Studies of the New Coronavirus 2019 (COVID-19): Reports from Saudi Arabia

Development of Serological Assays and Seroprevalence Studies of the New Coronavirus 2019 (COVID-19): Reports from Saudi Arabia

The Nuts and Bolts of SARS-CoV-2 Spike Receptor-Binding Domain Heterologous Expression

The SARS-CoV-2 spike residues 616/644 and 1138/1169 delineate two antibody epitopes in COVID-19 mRNA COMIRNATY vaccine (Pfizer/BioNTech)

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