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Evaluation of a novel multiplexed assay for determining IgG levels and functional activity to SARS-CoV-2

Marina Johnson, Helen R. Wagstaffe, Kimberly C. Gilmour, Annabelle Lea Mai, Joanna Lewis, Adam Hunt, Jake Sirr, Christopher Bengt, Louis Grandjean, View ORCID ProfileDavid Goldblatt
doi: https://doi.org/10.1101/2020.07.20.213249
Marina Johnson
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
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Helen R. Wagstaffe
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
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Kimberly C. Gilmour
bGreat Ormond Street Children’s Hospital NHS Foundation Trust, Great Ormond Street, London, WC1N 3JH, UK
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Annabelle Lea Mai
bGreat Ormond Street Children’s Hospital NHS Foundation Trust, Great Ormond Street, London, WC1N 3JH, UK
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Joanna Lewis
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
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Adam Hunt
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
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Jake Sirr
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
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Christopher Bengt
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
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Louis Grandjean
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
bGreat Ormond Street Children’s Hospital NHS Foundation Trust, Great Ormond Street, London, WC1N 3JH, UK
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David Goldblatt
aGreat Ormond Street Institute of Child Health, University College London, 30 Guilford Street, London, WC1N 1 EH, UK
bGreat Ormond Street Children’s Hospital NHS Foundation Trust, Great Ormond Street, London, WC1N 3JH, UK
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  • ORCID record for David Goldblatt
  • For correspondence: d.goldblatt@ucl.ac.uk
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Abstract

Background The emergence of SARS-CoV-2 has led to the development of new serological assays that could aid in diagnosis and evaluation of seroprevalence to inform an understanding of the burden of COVID-19 disease. Many available tests lack rigorous evaluation and therefore results may be misleading.

Objectives The aim of this study was to assess the performance of a novel multiplexed immunoassay for the simultaneous detection of antibodies against SARS-CoV-2 trimeric spike (S), spike receptor binding domain (RBD), spike N terminal domain and nucleocapsid antigen and a novel pseudo-neutralisation assay.

Methods A multiplexed solid-phase chemiluminescence assay (Meso Scale Discovery) was evaluated for the simultaneous detection of IgG binding to four SARS-CoV-2 antigens and the quantification of antibody-induced ACE-2 binding inhibition (pseudo-neutralisation assay). Sensitivity was evaluated with a total of 196 COVID-19 serum samples (169 confirmed PCR positive and 27 anti-nucleocapsid IgG positive) from individuals with mild symptomatic or asymptomatic disease. Specificity was evaluated with 194 control serum samples collected from adults prior to December 2019.

Results The specificity and sensitivity of the binding IgG assay was highest for S protein with a specificity of 97.4% and sensitivity of 96.2% for samples taken 14 days and 97.9% for samples taken 21 days following the onset of symptoms. IgG concentration to S and RBD correlated strongly with percentage inhibition measured by the pseudo-neutralisation assay.

Conclusion Excellent sensitivity for IgG detection was obtained over 14 days since onset of symptoms for three SARS-CoV-2 antigens (S, RBD and N) in this multiplexed assay which can also measure antibody functionality.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted July 21, 2020.
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Evaluation of a novel multiplexed assay for determining IgG levels and functional activity to SARS-CoV-2
Marina Johnson, Helen R. Wagstaffe, Kimberly C. Gilmour, Annabelle Lea Mai, Joanna Lewis, Adam Hunt, Jake Sirr, Christopher Bengt, Louis Grandjean, David Goldblatt
bioRxiv 2020.07.20.213249; doi: https://doi.org/10.1101/2020.07.20.213249
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Evaluation of a novel multiplexed assay for determining IgG levels and functional activity to SARS-CoV-2
Marina Johnson, Helen R. Wagstaffe, Kimberly C. Gilmour, Annabelle Lea Mai, Joanna Lewis, Adam Hunt, Jake Sirr, Christopher Bengt, Louis Grandjean, David Goldblatt
bioRxiv 2020.07.20.213249; doi: https://doi.org/10.1101/2020.07.20.213249

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