Going beyond clinical routine in SARS-CoV-2 antibody testing - A multiplex corona virus antibody test for the evaluation of cross-reactivity to endemic coronavirus antigens

Becker, Matthias; Strengert, Monika; Junker, Daniel; Kerrinnes, Tobias; Kaiser, Philipp; Traenkle, Bjoern; Dinter, Heiko; Haering, Julia; Zeck, Anne; Weise, Frank; Peter, Andreas; Hoerber, Sebastian; Fink, Simon; Ruoff, Felix; Bakchoul, Tamam; Baillot, Armin; Lohse, Stefan; Cornberg, Markus; Illig, Thomas; Gottlieb, Jens; Smola, Sigrun; Karch, André; Berger, Klaus; Rammensee, Hans‐Georg; Schenke‐Layland, Katja; Nelde, Annika; Maerklin, Melanie; Walz, Juliane S.; Templin, Markus F.; Joos, Thomas; Rothbauer, Ulrich; Krause, Gérard; Schneiderhan‐Marra, Nicole

doi:10.1101/2020.07.17.20156000

Cited by 12 publications

(19 citation statements)

References 21 publications

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“…Next, we analyzed whether the selected Nbs can block the interaction between the isolated receptor-binding domain (RBD), S1 domain (S1) or homotrimeric spike protein (spike) of SARS-CoV-2 to the human ACE2 receptor. To this end, we utilized a multiplex ACE2 competition assay employing the respective SARS-CoV-2 antigens coupled to paramagnetic beads (MagPlex Microspheres) 23 and incubated them with biotinylated ACE2 and dilutions of purified Nbs before measuring the residual binding of ACE2.…”

Section: Selection Of Sars-cov-2-specific Nbsmentioning

confidence: 99%

“…Currently available serological assays provide data on the presence and distribution of antibody subtypes against different SARS-CoV-2 antigens within serum samples of infected and recovered SARS-CoV-2 patients 4,17,22,23,26 . However, they do not differentiate between total and neutralizing RBD binding antibodies, which sterically inhibit viral entry via ACE2 7,8 .…”

Section: Neutrobodyplex -Using Nbs To Determine a Sars-cov-2 Neutralimentioning

confidence: 99%

“…We speculated that NM1267 specifically displaces such antibodies from the RBD:ACE2 interface, which can be monitored as a declining IgG signal (Figure 6 a). To test this hypothesis, we generated a high-throughput competitive binding assay, termed NeutrobodyPlex, by implementing the NM1267 in a recently developed, multiplexed serological assay 23 . We co-…”

Section: Neutrobodyplex -Using Nbs To Determine a Sars-cov-2 Neutralimentioning

confidence: 99%

“…Global fits were determined using the BLItzPro software and the global dissociation constant (KD) was calculated. Bound serum IgGs were detected via anti-human-IgG-PE as previously described 23 .…”

Section: Biophysical Biolayer Interferometry (Bli)mentioning

confidence: 99%

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NeutrobodyPlex - Nanobodies to monitor a SARS-CoV-2 neutralizing immune response

Wagner

Kaiser

Gramlich

et al. 2020

Preprint

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Facing the worldwide disease progression of COVID-19 caused by the SARS-CoV-2 virus, the situation is highly critical and there is an unmet need for effective vaccination, reliable diagnosis and therapeutic intervention. Neutralizing binding molecules such as antibodies or derivatives thereof have become important tools for acute treatment of COVID-19. Additionally, such binders provide the unique possibility to monitor the emergence and presence of a neutralizing immune response in infected or vaccinated individuals. Here we describe a set of 11 unique nanobodies (Nbs), originated from an immunized alpaca which bind with high affinities to the glycosylated SARS-CoV-2 Spike receptor domain (RBD). Using a multiplex in vitro binding assay we showed that eight of the selected Nbs effectively block the interaction between RBD, S1-domain and homotrimeric Spike protein with the angiotensin converting enzyme 2 (ACE2) as the viral docking site on human cells. According to competitive binding analysis and detailed epitope mapping, we grouped all Nbs blocking the RBD:ACE2 interaction in three distinct Nb-Sets and demonstrated their neutralizing effect with IC50 values in the low nanomolar range in a cell-based SARS-CoV-2 neutralization assay. Tested Nb combinations from different sets showed substantially lower IC50 values in both functional assays indicating a profound synergistic effect of Nbs simultaneously targeting different epitopes within the RBD. Finally, we applied the most potent Nb combinations in a competitive multiplex binding assay which we termed NeutrobodyPlex and detected a neutralizing immune response in plasma samples of infected individuals. We envisage that our Nbs have a high potential for prophylactic as well as therapeutic options and provide a novel approach to screen for a neutralizing immune response in infected or vaccinated individuals thus helping to monitor the immune status or to guide vaccine design.

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Section: Selection Of Sars-cov-2-specific Nbsmentioning

confidence: 99%

Section: Neutrobodyplex -Using Nbs To Determine a Sars-cov-2 Neutralimentioning

confidence: 99%

Section: Neutrobodyplex -Using Nbs To Determine a Sars-cov-2 Neutralimentioning

confidence: 99%

Section: Biophysical Biolayer Interferometry (Bli)mentioning

confidence: 99%

See 2 more Smart Citations

NeutrobodyPlex - Nanobodies to monitor a SARS-CoV-2 neutralizing immune response

Wagner

Kaiser

Gramlich

et al. 2020

Preprint

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“…With respect to specificity of the assay, the potential of cross-reactive antibody responses resulting from previous infections with other CoVs including SARS-CoV-1 and endemic ccCoVs NL63, HKU1, 229E and OC43, should be acknowledged (Becker et al, 2020;Meyer et al, 2014).…”

Section: Separate Detection Of Immunoglobulin M a And G Antibody Rementioning

confidence: 99%

From multiplex serology to serolomics – a novel approach to the antibody response against the SARS-CoV-2 proteome

Butt

Murugan

Hippchen

et al. 2020

Preprint

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Background: The emerging SARS-CoV-2 pandemic entails an urgent need for specific and sensitive high-throughput serological assays to assess SARS-CoV-2 epidemiology. We therefore aimed at developing a fluorescent-bead based SARS-CoV-2 multiplex serology assay for detection of antibody responses to the SARS-CoV-2 proteome. Methods: Proteins of the SARS-CoV-2 proteome and protein N of SARS-CoV-1 and common cold Coronaviruses (ccCoVs) were recombinantly expressed in E. coli or HEK293 cells. Assay performance was assessed in a Covid-19 case cohort (n=48 hospitalized patients from Heidelberg) as well as n=85 age- and sex-matched pre-pandemic controls from the ESTHER study. Assay validation included comparison with home-made immunofluorescence and commercial Enzyme-linked immunosorbent (ELISA) assays. Results: A sensitivity of 100% (95% CI: 86%-100%) was achieved in Covid-19 patients 14 days post symptom onset with dual sero-positivity to SARS-CoV-2 N and the receptor-binding domain of the spike protein. The specificity obtained with this algorithm was 100% (95% CI: 96%-100%). Antibody responses to ccCoVs N were abundantly high and did not correlate with those to SARS-CoV-2 N. Inclusion of additional SARS-CoV-2 proteins as well as separate assessment of immunoglobulin (Ig) classes M, A, and G allowed for explorative analyses regarding disease progression and course of antibody response. Conclusion: This newly developed SARS-CoV-2 multiplex serology assay achieved high sensitivity and specificity to determine SARS-CoV-2 sero-positivity. Its high throughput ability allows epidemiologic SARS-CoV-2 research in large population-based studies. Inclusion of additional pathogens into the panel as well as separate assessment of Ig isotypes will furthermore allow addressing research questions beyond SARS-CoV-2 sero-prevalence.

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Antibody tests for identification of current and past infection with SARS-CoV-2

Fox

Geppert

Dinnes

et al. 2022

Cochrane Database of Systematic Reviews

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Background The diagnostic challenges associated with the COVID‐19 pandemic resulted in rapid development of diagnostic test methods for detecting SARS‐CoV‐2 infection. Serology tests to detect the presence of antibodies to SARS‐CoV‐2 enable detection of past infection and may detect cases of SARS‐CoV‐2 infection that were missed by earlier diagnostic tests. Understanding the diagnostic accuracy of serology tests for SARS‐CoV‐2 infection may enable development of effective diagnostic and management pathways, inform public health management decisions and understanding of SARS‐CoV‐2 epidemiology. Objectives To assess the accuracy of antibody tests, firstly, to determine if a person presenting in the community, or in primary or secondary care has current SARS‐CoV‐2 infection according to time after onset of infection and, secondly, to determine if a person has previously been infected with SARS‐CoV‐2. Sources of heterogeneity investigated included: timing of test, test method, SARS‐CoV‐2 antigen used, test brand, and reference standard for non‐SARS‐CoV‐2 cases. Search methods The COVID‐19 Open Access Project living evidence database from the University of Bern (which includes daily updates from PubMed and Embase and preprints from medRxiv and bioRxiv) was searched on 30 September 2020. We included additional publications from the Evidence for Policy and Practice Information and Co‐ordinating Centre (EPPI‐Centre) ‘COVID‐19: Living map of the evidence’ and the Norwegian Institute of Public Health ’NIPH systematic and living map on COVID‐19 evidence’. We did not apply language restrictions. Selection criteria We included test accuracy studies of any design that evaluated commercially produced serology tests, targeting IgG, IgM, IgA alone, or in combination. Studies must have provided data for sensitivity, that could be allocated to a predefined time period after onset of symptoms, or after a positive RT‐PCR test. Small studies with fewer than 25 SARS‐CoV‐2 infection cases were excluded. We included any reference standard to define the presence or absence of SARS‐CoV‐2 (including reverse transcription polymerase chain reaction tests (RT‐PCR), clinical diagnostic criteria, and pre‐pandemic samples). Data collection and analysis We use standard screening procedures with three reviewers. Quality assessment (using the QUADAS‐2 tool) and numeric study results were extracted independently by two people. Other study characteristics were extracted by one reviewer and checked by a second. We present sensitivity and specificity with 95% confidence intervals (CIs) for each test and, for meta‐analysis, we fitted univariate random‐effects logistic regression models for sensitivity by eligible time period and for specificity by reference standard group. Heterogeneity was investigated by including indicator variables in the random‐effects logistic regression models. We tabulated result...

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Going beyond clinical routine in SARS-CoV-2 antibody testing - A multiplex corona virus antibody test for the evaluation of cross-reactivity to endemic coronavirus antigens

Cited by 12 publications

References 21 publications

NeutrobodyPlex - Nanobodies to monitor a SARS-CoV-2 neutralizing immune response

NeutrobodyPlex - Nanobodies to monitor a SARS-CoV-2 neutralizing immune response

From multiplex serology to serolomics – a novel approach to the antibody response against the SARS-CoV-2 proteome

Antibody tests for identification of current and past infection with SARS-CoV-2

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