Establishing SARS-CoV-2 membrane protein-specific antibodies as a valuable serological target via high-content microscopy

Daniel M Williams; Hailey R Hornsby; Ola M Shehata; Rebecca Brown; Marta Gallis; Naomi Meardon; Thomas A H Newman; Megan Plowright; Domen Zafred; Amber S M Shun-Shion; Anthony J Hodder; Deepa Bliss; Andrew Metcalfe; James R Edgar; David E Gordon; Jon R Sayers; Martin J Nicklin; Miles Carroll; PITCH Consortium; Paul J Collini; Stephen Brown; Thushan I de Silva; Andrew A Peden

doi:10.1016/j.isci.2023.107056

Establishing SARS-CoV-2 membrane protein-specific antibodies as a valuable serological target via high-content microscopy

iScience. 2023 Jul 21;26(7):107056. doi: 10.1016/j.isci.2023.107056. Epub 2023 Jun 7.

Authors

Daniel M Williams¹, Hailey R Hornsby², Ola M Shehata¹, Rebecca Brown², Marta Gallis², Naomi Meardon^{2

3}, Thomas A H Newman^{2

3}, Megan Plowright^{2

3}, Domen Zafred², Amber S M Shun-Shion¹, Anthony J Hodder¹, Deepa Bliss¹, Andrew Metcalfe¹, James R Edgar⁴, David E Gordon⁵, Jon R Sayers², Martin J Nicklin², Miles Carroll⁶; PITCH Consortium; Paul J Collini^{2

3}, Stephen Brown¹, Thushan I de Silva^{2

3}, Andrew A Peden¹

Affiliations

¹ School of Bioscience, University of Sheffield, Western Bank, Sheffield S10 2TN, UK.
² Department of Infection, Immunity and Cardiovascular Diseases, University of Sheffield Medical School, Beech Hill Road, Sheffield S10 2RX, UK.
³ South Yorkshire Regional Department of Infection and Tropical Medicine, Sheffield Teaching Hospitals NHS Foundation Trust, Glossop Road, Sheffield S10 2JF, UK.
⁴ Department of Pathology, University of Cambridge, Cambridge CB2 1QP, UK.
⁵ Department of Pathology, Emory University, Whitehead Building, Atlanta, GA, USA.
⁶ Wellcome Centre for Human Genetics, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7BN, UK.

Abstract

The prevalence and strength of serological responses mounted toward SARS-CoV-2 proteins other than nucleocapsid (N) and spike (S), which may be of use as additional serological markers, remains underexplored. Using high-content microscopy to assess antibody responses against full-length StrepTagged SARS-CoV-2 proteins, we found that 85% (166/196) of unvaccinated individuals with RT-PCR confirmed SARS-CoV-2 infections and 74% (31/42) of individuals infected after being vaccinated developed detectable IgG against the structural protein M, which is higher than previous estimates. Compared with N antibodies, M IgG displayed a shallower time-dependent decay and greater specificity. Sensitivity for SARS-CoV-2 seroprevalence was enhanced when N and M IgG detection was combined. These findings indicate that screening for M seroconversion may be a good approach for detecting additional vaccine breakthrough infections and highlight the potential to use HCM as a rapidly deployable method to identify the most immunogenic targets of newly emergent pathogens.

Keywords: Biological sciences; Cell biology; Immunology; Microbiology.

Grants and funding

WT_/Wellcome Trust/United Kingdom