Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay

Mariën, Joachim; Ceulemans, Ann; Míchiels, Johan; Heyndríckx, Leo; Kerkhof, Karen; Foqué, Nikki; Widdowson, Marc‐Alain; Mortgat, Laure; Duysburgh, Els; Desombere, Isabelle; Jansens, Hilde; Esbroeck, Marjan Van; Ariën, Kevin K.

doi:10.1016/j.jviromet.2020.114025

Cited by 109 publications

(132 citation statements)

References 32 publications

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“…During the course of this work, many reports of serological diagnostic assays, including those based upon the Luminex format (15)(16)(17)(18)(19)(20)(21)(22)(23), have been presented. Rather than developing an additional diagnostic platform, we emphasize here the utility of the Luminex system as a research tool -for monitoring changes in antibody responses over time, detecting antibody responses in different sample matrices and for rapid adaptation and evaluation of new antigen formulations.…”

Section: Discussionmentioning

confidence: 99%

A flexible, pan-species, multi-antigen platform for the detection and monitoring of SARS-CoV-2-specific antibody responses

Shen

Forgacs

Chapla

et al. 2021

Preprint

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The SARS-CoV-2 pandemic and the vaccination effort that is ongoing has created an unmet need for accessible, affordable, flexible and precise platforms for monitoring the induction, specificity and maintenance of virus-specific immune responses. Herein we validate a multiplex (Luminex-based) assay capable of detecting SARS-CoV-2-specific antibodies irrespective of host species, antibody isotype, and specimen type (e.g. plasma, serum, saliva or blood spots). The well-established precision of Luminex-based assays provides the ability to follow changes in antibody levels over time to many antigens, including multiple permutations of the most common SARS-CoV-2 antigens. This platform can easily measure antibodies known to correlate with neutralization activity as well as multiple non-SARS-CoV-2 antigens such as vaccines (e.g. Tetanus toxoid) or those from frequently encountered agents (influenza), which serve as stable reference points for quantifying the changing SARS-specific responses. All of the antigens utilized in our study can be made in-house, many in E. coli using readily available plasmids. Commercially sourced antigens may also be incorporated and newly available antigen variants can be rapidly produced and integrated, making the platform adaptable to the evolving viral strains in this pandemic.Brief SummaryA multi-antigen assay for monitoring SARS-CoV-2-specific antibodies irrespective of host species, antibody isotype, and specimen type was developed.

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Section: Discussionmentioning

confidence: 99%

A flexible, pan-species, multi-antigen platform for the detection and monitoring of SARS-CoV-2-specific antibody responses

Shen

Forgacs

Chapla

et al. 2021

Preprint

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“…A handful of microsphere-based SARS-CoV-2 serology assays have been developed, facilitating high throughput multiplex strategies for antibody detection (19)(20)(21)(22)(23). Multiplex for use under a CC0 license.…”

Section: Introductionmentioning

confidence: 99%

Antigen-based multiplex strategies to discriminate SARS-CoV-2 natural and vaccine induced immunity from seasonal human coronavirus humoral responses

Laing

et al. 2021

Preprint

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Sensitive and specific SARS-CoV-2 antibody assays remain critical for community and hospital-based SARS-CoV-2 sero-surveillance. With the rollout of SARS-CoV-2 vaccines, such assays must be able to distinguish vaccine from natural immunity to SARS-CoV-2 and related human coronaviruses. Here, we developed and implemented multiplex microsphere-based immunoassay strategies for COVD-19 antibody studies that incorporates spike protein trimers of SARS-CoV-2 and the endemic seasonal human coronaviruses (HCoV), enabling high throughout measurement of pre-existing cross-reactive antibodies. We varied SARS-CoV-2 antigen compositions within the multiplex assay, allowing direct comparisons of the effects of spike protein, receptor-binding domain protein (RBD) and nucleocapsid protein (NP) based SARS-CoV-2 antibody detection. Multiplex immunoassay performance characteristics are antigen-dependent, and sensitivities and specificities range 92-99% and 94-100%, respectively, for human subject samples collected as early as 7-10 days from symptom onset. SARS-CoV-2 spike and RBD had a strong correlative relationship for the detection of IgG. Correlation between detectable IgG reactive with spike and NP also had strong relationship, however, several PCR-positive and spike IgG-positive serum samples were NP IgG-negative. This spike and NP multiplex immunoassay has the potential to be useful for differentiation between vaccination and natural infection induced antibody responses. We also assessed the induction of de novo SARS-CoV-2 IgG cross reactions with SARS-CoV and MERS-CoV spike proteins. Furthermore, multiplex immunoassays that incorporate spike proteins of SARS-CoV-2 and HCoVs will permit investigations into the influence of HCoV antibodies on COVID-19 clinical outcomes and SARS-CoV-2 antibody durability.

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“…Samples were also screened by an in-house cVNT and Luminex MIA as reference, which are described in detail in Mariën et al 2020 (6). Briefly, for the cVNT, serial dilutions of serum (1/50-1/1600) were incubated with 3xTCID100 of a primary isolate of SARS-CoV-2 during 1 h. This solution was added to Vero cells (18.000cells/well) in a 96well plate and incubated for 5 days (37 •C / 7 % CO2).…”

Section: Methodsmentioning

confidence: 99%

“…Samples were still considered to be positive if more than 10% reactivity was observed at a 1/50 serum dilution. For the Luminex MIA, recombinant receptor binding domain (RBD) and Nucleocapsid protein (NCP) (BIOCONNECT, The Netherlands) were coupled to 1.25x10 6 paramagnetic MAGPLEX COOH-microspheres from Luminex Corporation (Texas, USA). After incubation of beads and diluted sera (1/300), a biotin-labelled anti-human IgG (1:125) and streptavidin-R-phycoerythrin (1:1000) conjugate was added.…”

Section: Methodsmentioning

confidence: 99%

“…While a plethora of serological tests became available months after the emergence of SARS-CoV-2, not all of them are appropriate for large-scale serosurveillance. Most high-throughput tests detect total BAbs only, such as enzyme-linked immunosorbent (ELISA), lateral flow (LFA) or multiplex (MIA) immunoassays (6). These tests can be run in basic diagnostic labs and are mainly used to confirm past infection, but are unable to directly show the presence of neutralizing antibodies in serum.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of a surrogate virus neutralization test for high-throughput serosurveillance of SARS-CoV-2

Mariën

Míchiels

Heyndríckx

et al. 2021

Preprint

Self Cite

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High-throughput serological tests that can detect neutralizing antibodies against SARS-CoV-2 are desirable for serosurveillance and vaccine efficacy evaluation. Although the conventional neutralization test (cVNT) remains the gold standard to confirm the presence of neutralizing antibodies in sera, the test is too labour-intensive for massive screening programs and less reproducible as live virus and cell culture is involved. Here, we performed an independent evaluation of a commercially available surrogate virus neutralization test (sVNT, GenScript cPass) that can be done without biosafety level 3 containment in less than 2 hours. When using the cVNT and a Luminex multiplex immunoassay (MIA) as reference, the sVNT obtained a sensitivity of 94% (CI 90-96%) on a panel of 317 immune sera that were obtained from hospitalized and mild COVID-19 cases from Belgium and a sensitivity of 89% (CI 81-93%) on a panel of 184 healthcare workers from the Democratic Republic of Congo. We also found strong antibody titer correlations (rs>0.8) among the different techniques used. In conclusion, our evaluation suggests that the sVNT could be a powerful tool to monitor/detect neutralising antibodies in cohort and population studies. The technique could be especially useful for vaccine evaluation studies in sub-Saharan Africa where the basic infrastructure to perform cVNTs is lacking.

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Evaluating SARS-CoV-2 spike and nucleocapsid proteins as targets for antibody detection in severe and mild COVID-19 cases using a Luminex bead-based assay

Cited by 109 publications

References 32 publications

A flexible, pan-species, multi-antigen platform for the detection and monitoring of SARS-CoV-2-specific antibody responses

A flexible, pan-species, multi-antigen platform for the detection and monitoring of SARS-CoV-2-specific antibody responses

Antigen-based multiplex strategies to discriminate SARS-CoV-2 natural and vaccine induced immunity from seasonal human coronavirus humoral responses

Evaluation of a surrogate virus neutralization test for high-throughput serosurveillance of SARS-CoV-2

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