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Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva

Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva
Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva

Detecting antibody responses during and after SARS-CoV-2 infection is essential in determining the seroepidemiology of the virus and the potential role of antibody in disease. Scalable, sensitive and specific serological assays are essential to this process. The detection of antibody in hospitalized patients with severe disease has proven relatively straightforward; detecting responses in subjects with mild disease and asymptomatic infections has proven less reliable. We hypothesized that the suboptimal sensitivity of antibody assays and the compartmentalization of the antibody response may contribute to this effect. We systematically developed an ELISA, optimizing different antigens and amplification steps, in serum and saliva from non-hospitalized SARS-CoV-2-infected subjects. Using trimeric spike glycoprotein, rather than nucleocapsid, enabled detection of responses in individuals with low antibody responses. IgG1 and IgG3 predominate to both antigens, but more anti-spike IgG1 than IgG3 was detectable. All antigens were effective for detecting responses in hospitalized patients. Anti-spike IgG, IgA and IgM antibody responses were readily detectable in saliva from a minority of RT-PCR confirmed, non-hospitalized symptomatic individuals, and these were mostly subjects who had the highest levels of anti-spike serum antibodies. Therefore, detecting antibody responses in both saliva and serum can contribute to determining virus exposure and understanding immune responses after SARS-CoV-2 infection.

COVID-19, ELISA, SARS-CoV-2, antibodies
0019-2805
135-147
Faustini, Sian E
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Jossi, Sian E
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Perez-Toledo, Marisol
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Shields, Adrian M
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Allen, Joel D
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Watanabe, Yasunori
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Newby, Maddy L
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Cook, Alex
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Willcox, Carrie R
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Salim, Mahboob
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Goodall, Margaret
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Heaney, Jennifer L
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Marcial-Juarez, Edith
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Morley, Gabriella L
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Torlinska, Barbara
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Wraith, David C
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Veenith, Tonny V
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Harding, Stephen
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Jolles, Stephen
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Ponsford, Mark J
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Plant, Tim
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Huissoon, Aarnoud
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O'Shea, Matthew K
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Willcox, Benjamin E
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Drayson, Mark T
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Crispin, Max
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Cunningham, Adam F
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Richter, Alex G
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Faustini, Sian E
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Jossi, Sian E
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Perez-Toledo, Marisol
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Shields, Adrian M
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Allen, Joel D
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Watanabe, Yasunori
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Newby, Maddy L
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Cook, Alex
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Willcox, Carrie R
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Salim, Mahboob
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Goodall, Margaret
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Heaney, Jennifer L
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Marcial-Juarez, Edith
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Morley, Gabriella L
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Torlinska, Barbara
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Wraith, David C
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Veenith, Tonny V
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Harding, Stephen
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Jolles, Stephen
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Ponsford, Mark J
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Plant, Tim
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Huissoon, Aarnoud
c2d44c09-b103-4be0-8e71-10ed90892b39
O'Shea, Matthew K
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Willcox, Benjamin E
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Drayson, Mark T
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Crispin, Max
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Cunningham, Adam F
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Richter, Alex G
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Faustini, Sian E, Jossi, Sian E, Perez-Toledo, Marisol, Shields, Adrian M, Allen, Joel D, Watanabe, Yasunori, Newby, Maddy L, Cook, Alex, Willcox, Carrie R, Salim, Mahboob, Goodall, Margaret, Heaney, Jennifer L, Marcial-Juarez, Edith, Morley, Gabriella L, Torlinska, Barbara, Wraith, David C, Veenith, Tonny V, Harding, Stephen, Jolles, Stephen, Ponsford, Mark J, Plant, Tim, Huissoon, Aarnoud, O'Shea, Matthew K, Willcox, Benjamin E, Drayson, Mark T, Crispin, Max, Cunningham, Adam F and Richter, Alex G (2021) Development of a high-sensitivity ELISA detecting IgG, IgA and IgM antibodies to the SARS-CoV-2 spike glycoprotein in serum and saliva. Immunology, 164 (1), 135-147. (doi:10.1111/imm.13349).

Record type: Article

Abstract

Detecting antibody responses during and after SARS-CoV-2 infection is essential in determining the seroepidemiology of the virus and the potential role of antibody in disease. Scalable, sensitive and specific serological assays are essential to this process. The detection of antibody in hospitalized patients with severe disease has proven relatively straightforward; detecting responses in subjects with mild disease and asymptomatic infections has proven less reliable. We hypothesized that the suboptimal sensitivity of antibody assays and the compartmentalization of the antibody response may contribute to this effect. We systematically developed an ELISA, optimizing different antigens and amplification steps, in serum and saliva from non-hospitalized SARS-CoV-2-infected subjects. Using trimeric spike glycoprotein, rather than nucleocapsid, enabled detection of responses in individuals with low antibody responses. IgG1 and IgG3 predominate to both antigens, but more anti-spike IgG1 than IgG3 was detectable. All antigens were effective for detecting responses in hospitalized patients. Anti-spike IgG, IgA and IgM antibody responses were readily detectable in saliva from a minority of RT-PCR confirmed, non-hospitalized symptomatic individuals, and these were mostly subjects who had the highest levels of anti-spike serum antibodies. Therefore, detecting antibody responses in both saliva and serum can contribute to determining virus exposure and understanding immune responses after SARS-CoV-2 infection.

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Accepted/In Press date: 14 April 2021
e-pub ahead of print date: 1 May 2021
Published date: September 2021
Additional Information: Research funding Global Challenges Research Fund UK National Institute for Health Research, Birmingham Biomedical Research Centres Funding Scheme Welsh Clinical Academic Training University of Birmingham Scripps Consortium for HIV Vaccine Development (NIH: Nation Institute for Allergy and Infectious Diseases). Grant Number: AI144462 NIH Graduate Partnership Program The Medical Research Council International AIDS Vaccine Initiative Collaboration for AIDS Vaccine Discovery. Grant Numbers: OPP1084519, OPP1115782, OPP1196345/INV-008813 The Institute for Global Innovation. Grant Number: 3107 University of Southampton Coronavirus Response Fund University Hospitals Birmingham NHS Foundation Trust ACKNOWLEDGMENTS We would like to thank the University of Birmingham Clinical Immunology Service for their invaluable support in sample collection and processing. We would also like to thank the National Institute for Health Research (NIHR)/Wellcome Trust Birmingham Clinical Research Facility staff for their vital support in study participant recruitment and sample collection. We also gratefully acknowledge the University of Birmingham Protein Expression Facility for use of mammalian expression equipment. The CoCo study was carried out at the National Institute for Health Research (NIHR)/Wellcome Trust Birmingham Clinical Research Facility. The views expressed are those of the authors(s) and not necessarily those of the NHS, the NIHR or the Department of Health.
Keywords: COVID-19, ELISA, SARS-CoV-2, antibodies

Identifiers

Local EPrints ID: 449804
URI: http://eprints.soton.ac.uk/id/eprint/449804
ISSN: 0019-2805
PURE UUID: b1200902-b594-42a2-8f93-1d414316f042
ORCID for Joel D Allen: ORCID iD orcid.org/0000-0003-2547-968X
ORCID for Max Crispin: ORCID iD orcid.org/0000-0002-1072-2694

Catalogue record

Date deposited: 17 Jun 2021 16:36
Last modified: 17 Mar 2024 04:09

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Contributors

Author: Sian E Faustini
Author: Sian E Jossi
Author: Marisol Perez-Toledo
Author: Adrian M Shields
Author: Joel D Allen ORCID iD
Author: Yasunori Watanabe
Author: Maddy L Newby
Author: Alex Cook
Author: Carrie R Willcox
Author: Mahboob Salim
Author: Margaret Goodall
Author: Jennifer L Heaney
Author: Edith Marcial-Juarez
Author: Gabriella L Morley
Author: Barbara Torlinska
Author: David C Wraith
Author: Tonny V Veenith
Author: Stephen Harding
Author: Stephen Jolles
Author: Mark J Ponsford
Author: Tim Plant
Author: Aarnoud Huissoon
Author: Matthew K O'Shea
Author: Benjamin E Willcox
Author: Mark T Drayson
Author: Max Crispin ORCID iD
Author: Adam F Cunningham
Author: Alex G Richter

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