SARS-CoV-2 Multi-Antigen Serology Assay.

Ramin Mazhari,Catherine Chen,Shazia Ruybal-Pesántez,Nicholas Kiernan-Walker,Deborah A Williamson,Stephanie Hyslop,Emily M Eriksson,Rhea J Longley,Leanne J Robinson,Fiona Angrisano,Caitlin Bourke,Ivo Mueller

doi:10.3390/mps4040072

Abstract

Serology tests are extremely useful for assessing whether a person has been infected with a pathogen. Since the onset of the COVID-19 pandemic, measurement of anti-SARS-CoV-2-specific antibodies has been considered an essential tool in identifying seropositive individuals and thereby understanding the extent of transmission in communities. The Luminex system is a bead-based technology that has the capacity to assess multiple antigens simultaneously using very low sample volumes and is ideal for high-throughput studies. We have adapted this technology to develop a COVID-19 multi-antigen serological assay. This protocol described here carefully outlines recommended steps to optimize and establish this method for COVID-19-specific antibody measurement in plasma and in saliva. However, the protocol can easily be customized and thus the assay is broadly applicable to measure antibodies to other pathogens.

Highlights

The pathogen causing coronavirus disease 2019 (COVID-19) has been identified as a novel zoonotic coronavirus termed the severe acute respiratory syndrome coronavirus (SARS-CoV-2)
Production of recombinant proteins commonly uses a range of expression systems, including E. coli, HEK cells and Baculovirus system
It is well established that the glycosylation capabilities of different species differ significantly, resulting in varying glycosylation between recombinant proteins expressed in mammalian, yeast and insect cells [22]

Summary

Introduction

The pathogen causing coronavirus disease 2019 (COVID-19) has been identified as a novel zoonotic coronavirus termed the severe acute respiratory syndrome coronavirus (SARS-CoV-2). IgG and IgM anti-SARS-CoV-2 antibodies are generated in most infected individuals, though mild and asymptomatic infections result in relatively low antibody levels [1]. Kinetic studies to date suggest that both IgM and IgG anti-SARS-CoV-2 antibodies are detectable approximately 7–10 days after the onset of clinical symptoms, the longevity of IgM is shorter than IgG [2]. Longitudinal studies have demonstrated the presence of antibodies and the neutralizing effects of these antibodies after 8–10 months in people who have recovered from both mild and severe COVID-19 disease [3,4]. There have been indications that pre-existing antibodies to seasonal coronaviruses may confer some level of protective immunity to severe COVID-19 disease [6,7]

Methods

Results

Conclusion