Evaluation of S- and M-Proteins Expressed in IEscherichia coli/I and HEK Cells for Serological Detection of Antibodies in Response to SARS-CoV-2 Infections and mRNA-Based Vaccinations

• Published in: Pathogens (Basel) Vol. 11; no. 12
• Main Authors: Schwarze, Mandy, Luo, Ji, Brakel, Alexandra, Krizsan, Andor, Lakowa, Nicole, Grünewald, Thomas, Lehmann, Claudia, Wolf, Johannes, Borte, Stephan, Milkovska-Stamenova, Sanja, Gabert, Jörg, Scholz, Markus, Hoffmann, Ralf
• Format: Journal Article
• Language: English
• Published: MDPI AG 01.12.2022
• Subjects: Bacterial proteins; Enzyme-linked immunosorbent assay; Escherichia coli; Immune response; Immune system; Immunological research; Physiological aspects; Testing; Germany
• ISSN: 2076-0817; 2076-0817
• DOI: 10.3390/pathogens11121515

This study investigated the IgG and IgA antibody response against recombinant S1 and receptor binding domains (RBD) of the spike (S-) protein and the membrane (M-) protein using a set of 115 serum samples collected from patients infected with SARS-CoV-2 in Germany before April 2021 using protein and peptide ELISA. As S1- and RBD-proteins expressed in Escherichia coli provided poor sensitivities in ELISA, they were replaced by proteins expressed in HEK cells. The RBD-ELISA provided a sensitivity of 90.6% (N = 85) for samples collected from patients with confirmed SARS-CoV-2 infections more than 14 days after symptom onset or a positive PCR test. In population-based controls, the specificity was 97.9% (N = 94). In contrast, the sensitivities were only 41.2% and 72.6% for M- and N-proteins, respectively, while the specificities were 88.5% and 100%, respectively. Considering also 20 samples collected during the first two weeks of symptom onset or PCR confirmation, the sensitivity of RBD- and N-protein ELISA decreased to 82.6% and 72.6%, respectively. The combination of two data sets, i.e., N- and RBD-, N- and M-, or RBD- and M-proteins increased the sensitivity to 85.8%, 77.9%, and 87.8%, respectively. Peptide mapping mostly confirmed epitopes previously reported for S1- and M-proteins, but they were only recognized by a few samples already tested positive in the corresponding protein ELISA indicating that peptide-based assays will not improve the diagnostic sensitivity.