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. 2022 May 16;12(1):8067.
doi: 10.1038/s41598-022-10823-7.

Development of a new antigen-based microarray platform for screening and detection of human IgG antibodies against SARS-CoV-2

Sindy Burgold-Voigt  1   2 , Elke Müller  3   4 , David Zopf  3   5 , Stefan Monecke  3   4   6 , Sascha D Braun  3   4 , Katrin Frankenfeld  7 , Michael Kiehntopf  8 , Sebastian Weis  9   10 , Thomas Schumacher  11 , Mathias W Pletz  9 , Ralf Ehricht  3   4   5 ; CoNAN Study Group
Collaborators, Affiliations

Development of a new antigen-based microarray platform for screening and detection of human IgG antibodies against SARS-CoV-2

Sindy Burgold-Voigt et al. Sci Rep. .

Abstract

Strategies to contain the current SARS-CoV-2 pandemic rely, beside vaccinations, also on molecular and serological testing. For any kind of assay development, screening for the optimal antigen is essential. Here we describe the verification of a new protein microarray with different commercially available preparations significant antigens of SARS-CoV-2 that can be used for the evaluation of the performance of these antigens in serological assays and for antibody screening in serum samples. Antigens of other pathogens that are addressed by widely used vaccinations were also included. To evaluate the accuracy of 21 different antigens or antigen preparations on the microarray, receiver operating characteristics (ROC) curve analysis using ELISA results as reference were performed. Except for a single concentration, a diagnostic sensitivity of 1 was determined for all antigen preparations. A diagnostic specificity, as well as an area under the curve (AUC) of 1 was obtained for 16 of 21 antigen preparations. For the remaining five, the diagnostic specificity ranged from 0.942 to 0.981 and AUC from 0.974 to 0.999. The optimized assay was subsequently also applied to determine the immune status of previously tested individuals and/or to detect the immunization status after COVID-19 vaccination. Microarray evaluation of the antibody profiles of COVID-19 convalescent and post vaccination sera showed that the IgG response differed between these groups, and that the choice of the test antigen is crucial for the assay performance. Furthermore, the results showed that the immune response is highly individualized, depended on several factors (e.g., age or sex), and was not directly related to the severity of disease. The new protein microarray provides an ideal method for the parallel screening of many different antigens of vaccine-preventable diseases in a single sample and for reliable and meaningful diagnostic tests, as well as for the development of safe and specific vaccines.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Verification of the new SARS-CoV2-VAC-antigen microarray: 101 ELISA-tested sera from the CoNAN study were tested with a total of 14 antigens spotted at different concentrations and in different buffer conditions.
Figure 2
Figure 2
Verification of the additional vaccination antigens on microarray: 24 ELISA-tested sera were tested on the new microarray, 3 typical vaccine antigens were evaluated and used for further analyses.
Figure 3
Figure 3
Shown are the results of the new SARS-CoV2-VAC antigen microarray and the available results of real-time (RT)-PCR, lateral flow (LF) and ELISA tests for a total of 112 sera tested, with some subjects tested multiple times, at different time points.
Figure 4
Figure 4
The SARS-CoV2-VAC antigen microarray was used to investigate immunization status after COVID-19 vaccination. Shown are the results of 35 subjects tested who received BioNTech/Pfizer's Cormirnaty vaccine and 8 subjects tested who received AstraZeneca's Vaxzevria vaccine. Sera taken prior vaccination.
Figure 5
Figure 5
The verification of the new SARS-CoV2-VAC microarray was carried out according to the steps indicated in the scheme.
See this image and copyright information in PMC

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