Profiling of linear B-cell epitopes against human coronaviruses in pooled sera sampled early in the COVID-19 pandemic

Background: Antibodies play a key role in the immune defence against infectious pathogens. Understanding the underlying process of B cell recognition is not only of fundamental interest; it supports important applications within diagnostics and therapeutics. Whereas the nature of conformational B cell epitope recognition is inherently complicated, linear B cell epitopes offer a straightforward approach that potentially can be reduced to one of peptide recognition.
Methods: Using an overlapping peptide approach representing the entire proteomes of the seven main coronaviruses known to infect humans, we analysed sera pooled from eight PCR-confirmed COVID-19 convalescents and eight pre-pandemic controls. Using a high-density peptide microarray platform, 13-mer peptides overlapping by 11 amino acids were in situ synthesised and incubated with the pooled primary serum samples, followed by development with secondary fluorochrome-labelled anti-IgG and -IgA antibodies. Interactions were detecte..., Peptide microarray design
Peptide microarrays were designed using the proteomes of the seven human coronaviruses (HcoVs):
·      HCoV-229E: 7 proteins
·      HCoV-HKU1(N1): 8 proteins
·      HCoV-NL63: 6 proteins
·      HCoV-OC43: 9 proteins
·      MERS-CoV: 9 proteins
·      SARS-CoV: 14 proteins
·      SARS-CoV-2: 13 proteins
The open reading frame (ORF) 1a was excluded from all the HCoV proteomes since the ORF1ab covered these sequences. As a positive control pathogen, the entire proteome of the human cytomegalovirus (HCMV, strain AD169), consisting of 190 proteins, was included together with the entire proteome of the Zaire Ebola virus (strain Mayinga-76, EBOZM), consisting of 9 proteins. These 265 protein sequences were represented as 13 amino acid long peptides, overlapping by 11 amino acids and tiling by 2 amino acids. Leading to total of 66581 non-redundant virus-derived peptide sequences. As a source of background-binding control, 3900 non-overlapping 13 amino acid peptide sequ..., , # Data from: Profiling of linear B-cell epitopes against human coronaviruses in pooled sera sampled early in the COVID-19 pandemic

## [https://doi.org/10.5061/dryad.s1rn8pkg7](https://doi.org/10.5061/dryad.s1rn8pkg7)

The study used high-density peptide microarrays to investigate linear B cell epitopes in the seven human coronaviruses, human cytomegalovirus (strain AD169), and Zaire Ebola virus (strain Mayinga-76). Two serum pools derived from (1) eight convalescent PCR-confirmed COVID-19 patients and (2) eight individuals sampled prior to the COVID-19 pandemic (pre-pandemic) were analysed for seroreactivity to peptides derived from the nine viruses represented on the microarrays. Antibody responses from the pandemic serum pool serum were skewed towards recognising SARS-CoV-2-derived peptides, whereas the pre-pandemic serum pool was skewed towards recognising the common cold coronavirus (CCC) derived peptides. The study identified 333 human coronavirus derived linear B cell epitopes, w...