Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2

Alice Halliday; Anna E Long; Holly E Baum; Amy Thomas; Kathryn L Shelley; Elizabeth H Oliver; Kapil Gupta; Ore Francis; Maia Kavanagh Williamson; Natalie D Di Bartolo; Matthew J Randell; Yassin Ben Khoud; Ilana Kelland; Georgina L M Mortimer; Olivia E Ball; Charlie Plumptre; M A Chandler; Ulrike Obst; Massimiliano  Secchi; Lorenzo Piemonti; Vito Lampasona; Joyce Smith; Michaela Gregorova; Lea Knezevic; Jane Metz; Rachael S Barr; M B Morales-Aza; Jennifer L Oliver; Lucy Collingwood; Benjamin E Hitchings; Susan M Ring; Linda Wooldridge; Laura Rivino; Nicholas John  Timpson; Jorgen McKernon; Peter Muir; Fergus W Hamilton; David T Arnold; Dek N Woolfson; Anu Goenka; Andrew D Davidson; Ash M Toye; Imre Berger; Mick Bailey; Kathleen M Gillespie; Alistair J K Williams; Adam Finn

doi:10.3389/fimmu.2022.968317

Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2

Alice Halliday^*, Anna E Long, Holly E Baum, Amy Thomas, Kathryn L Shelley, Elizabeth H Oliver, Kapil Gupta, Ore Francis, Maia Kavanagh Williamson, Natalie D Di Bartolo, Matthew J Randell, Yassin Ben Khoud, Ilana Kelland, Georgina L M Mortimer, Olivia E Ball, Charlie Plumptre, M A Chandler, Ulrike Obst, Massimiliano Secchi, Lorenzo PiemontiVito Lampasona, Joyce Smith, Michaela Gregorova, Lea Knezevic, Jane Metz, Rachael S Barr, M B Morales-Aza, Jennifer L Oliver, Lucy Collingwood, Benjamin E Hitchings, Susan M Ring, Linda Wooldridge, Laura Rivino, Nicholas John Timpson, Jorgen McKernon, Peter Muir, Fergus W Hamilton, David T Arnold, Dek N Woolfson, Anu Goenka, Andrew D Davidson, Ash M Toye, Imre Berger, Mick Bailey, Kathleen M Gillespie, Alistair J K Williams, Adam Finn

^*Corresponding author for this work

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Abstract

Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97% specificity and >92% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.

Original language	English
Article number	968317
Number of pages	20
Journal	Frontiers in Immunology
Volume	13
DOIs	https://doi.org/10.3389/fimmu.2022.968317
Publication status	Published - 9 Nov 2022

Bibliographical note

Funding Information:
This work was supported by multiple grants to AH, AL, OF, AMT, and IB awarded by the Elizabeth Blackwell Institute, and funded in part by the Wellcome Trust [Grant number 204813/Z/16/Z] with additional support from Bristol Alumni and Friends. AL is funded by a Diabetes UK/JDRF RD Lawrence Fellowship (18/0005778 and 3-APF-2018-591-A-N). The DISCOVER study was supported by donations to Southmead Hospital Charity (Registered Charity Number: 1055900). The LOGIC study was funded by The Grand Appeal (The Official Bristol Children’s Hospital Charity; a registered charity in England and Wales (1043603)) through a grant awarded to AF and AG. ACT is supported by the Wellcome Trust (217509/Z/19/Z) and UKRI through the JUNIPER consortium MR/V038613/1 and CoMMinS study MR/V028545/1. The UK Medical Research Council and Wellcome (Grant ref: 217065/Z/19/Z) and the University of Bristol provide core support for ALSPAC. NT is a Wellcome Trust Investigator (202802/Z/16/Z), is the PI of the Avon Longitudinal Study of Parents and Children (MRC & WT 217065/Z/19/Z), is supported by the University of Bristol NIHR Biomedical Research Centre (BRC-1215-2001), the MRC Integrative Epidemiology Unit (MC_UU_00011/1) and works within the CRUK Integrative Cancer Epidemiology Programme (C18281/A29019). LIPS assay development was supported by a joint grant from Diabetes UK/JDRF (20/0006217) to KMG. IB is supported by the Wellcome Trust (106115/Z/14/Z, 221708/Z/20/Z), the ERC (contr. nrs. 834631, 963992) and the EPSRC Impact Acceleration Account EP/R511663/1. We also acknowledge funding from BBSRC/EPSRC Synthetic Biology Research Centre (BB/L01386X/1, to DNW, NB and AMT), NHS Blood and Transplant (WP15-05, to NB and AMT), and the NIHR Blood and Transplant Research Unit in Red Cell Products (IS-BTU-1214-10032, to NB and AMT). This publication is the work of the authors and AH et al. will serve as guarantors for the contents of this paper.

Funding Information:
AF is a member of the Joint Committee on Vaccination and Immunisation, the UK National Immunisation Technical Advisory Group and is chair of the WHO European Regional Technical Advisory Group of Experts (ETAGE) on immunization and ex officio a member of the WHO SAGE working group on COVID vaccines. He is investigator COVID-19 vaccine on studies and trials funded by Pfizer, Sanofi, Valneva, the Gates Foundation and the UK government. This manuscript presents independent research funded in part by the National Institute for Health Research (NIHR). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health and Social Care.

Publisher Copyright:
Copyright © 2022 Halliday, Long, Baum, Thomas, Shelley, Oliver, Gupta, Francis, Williamson, Di Bartolo, Randell, Ben-Khoud, Kelland, Mortimer, Ball, Plumptre, Chandler, Obst, Secchi, Piemonti, Lampasona, Smith, Gregorova, Knezevic, Metz, Barr, Morales-Aza, Oliver, Collingwood, Hitchings, Ring, Wooldridge, Rivino, Timpson, McKernon, Muir, Hamilton, Arnold, Woolfson, Goenka, Davidson, Toye, Berger, Bailey, Gillespie, Williams and Finn.

Structured keywords

Bristol BioDesign Institute
BrisSynBio

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.3389/fimmu.2022.968317Licence: CC BY

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This is the final published version of the article (version of record). It first appeared online via Frontiers Media at https://doi.org/10.3389/fimmu.2022.968317 .Please refer to any applicable terms of use of the publisher.
Final published version, 6.52 MBLicence: CC BY

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Elizabeth Blackwell Institute Early Career Fellowship
Halliday, A. & Finn, A. H. R.
1/10/18 → 30/09/20
Project: Research

Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2
Oliver, J. (Creator), Oliver, E. (Creator), Thomas, A. (Creator), Halliday, A. (Creator), Goenka, A. (Creator), Baum, H. (Creator), Gillespie, K. M. (Creator), Long, A. E. (Creator), Toye, A. M. (Creator) & Finn, A. H. R. (Creator), University of Bristol, 28 Sept 2022
DOI: 10.5523/bris.3bo7kz80s21fy2oknckvi8xq18, http://data.bris.ac.uk/data/dataset/3bo7kz80s21fy2oknckvi8xq18
Dataset

Cite this

Halliday, A., Long, A. E., Baum, H. E., Thomas, A., Shelley, K. L., Oliver, E. H., Gupta, K., Francis, O., Kavanagh Williamson, M., Di Bartolo, N. D., Randell, M. J., Ben Khoud, Y., Kelland, I., Mortimer, G. L. M., Ball, O. E., Plumptre, C., Chandler, M. A., Obst, U., Secchi, M., ... Finn, A. (2022). Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2. Frontiers in Immunology, 13, Article 968317. https://doi.org/10.3389/fimmu.2022.968317

@article{379110c308e6432c91615b397d81d92d,

title = "Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2",

abstract = "Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97% specificity and >92% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.",

author = "Alice Halliday and Long, {Anna E} and Baum, {Holly E} and Amy Thomas and Shelley, {Kathryn L} and Oliver, {Elizabeth H} and Kapil Gupta and Ore Francis and {Kavanagh Williamson}, Maia and {Di Bartolo}, {Natalie D} and Randell, {Matthew J} and {Ben Khoud}, Yassin and Ilana Kelland and Mortimer, {Georgina L M} and Ball, {Olivia E} and Charlie Plumptre and Chandler, {M A} and Ulrike Obst and Massimiliano Secchi and Lorenzo Piemonti and Vito Lampasona and Joyce Smith and Michaela Gregorova and Lea Knezevic and Jane Metz and Barr, {Rachael S} and Morales-Aza, {M B} and Oliver, {Jennifer L} and Lucy Collingwood and Hitchings, {Benjamin E} and Ring, {Susan M} and Linda Wooldridge and Laura Rivino and Timpson, {Nicholas John} and Jorgen McKernon and Peter Muir and Hamilton, {Fergus W} and Arnold, {David T} and Woolfson, {Dek N} and Anu Goenka and Davidson, {Andrew D} and Toye, {Ash M} and Imre Berger and Mick Bailey and Gillespie, {Kathleen M} and Williams, {Alistair J K} and Adam Finn",

note = "Funding Information: This work was supported by multiple grants to AH, AL, OF, AMT, and IB awarded by the Elizabeth Blackwell Institute, and funded in part by the Wellcome Trust [Grant number 204813/Z/16/Z] with additional support from Bristol Alumni and Friends. AL is funded by a Diabetes UK/JDRF RD Lawrence Fellowship (18/0005778 and 3-APF-2018-591-A-N). The DISCOVER study was supported by donations to Southmead Hospital Charity (Registered Charity Number: 1055900). The LOGIC study was funded by The Grand Appeal (The Official Bristol Children{\textquoteright}s Hospital Charity; a registered charity in England and Wales (1043603)) through a grant awarded to AF and AG. ACT is supported by the Wellcome Trust (217509/Z/19/Z) and UKRI through the JUNIPER consortium MR/V038613/1 and CoMMinS study MR/V028545/1. The UK Medical Research Council and Wellcome (Grant ref: 217065/Z/19/Z) and the University of Bristol provide core support for ALSPAC. NT is a Wellcome Trust Investigator (202802/Z/16/Z), is the PI of the Avon Longitudinal Study of Parents and Children (MRC & WT 217065/Z/19/Z), is supported by the University of Bristol NIHR Biomedical Research Centre (BRC-1215-2001), the MRC Integrative Epidemiology Unit (MC_UU_00011/1) and works within the CRUK Integrative Cancer Epidemiology Programme (C18281/A29019). LIPS assay development was supported by a joint grant from Diabetes UK/JDRF (20/0006217) to KMG. IB is supported by the Wellcome Trust (106115/Z/14/Z, 221708/Z/20/Z), the ERC (contr. nrs. 834631, 963992) and the EPSRC Impact Acceleration Account EP/R511663/1. We also acknowledge funding from BBSRC/EPSRC Synthetic Biology Research Centre (BB/L01386X/1, to DNW, NB and AMT), NHS Blood and Transplant (WP15-05, to NB and AMT), and the NIHR Blood and Transplant Research Unit in Red Cell Products (IS-BTU-1214-10032, to NB and AMT). This publication is the work of the authors and AH et al. will serve as guarantors for the contents of this paper. Funding Information: AF is a member of the Joint Committee on Vaccination and Immunisation, the UK National Immunisation Technical Advisory Group and is chair of the WHO European Regional Technical Advisory Group of Experts (ETAGE) on immunization and ex officio a member of the WHO SAGE working group on COVID vaccines. He is investigator COVID-19 vaccine on studies and trials funded by Pfizer, Sanofi, Valneva, the Gates Foundation and the UK government. This manuscript presents independent research funded in part by the National Institute for Health Research (NIHR). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health and Social Care. Publisher Copyright: Copyright {\textcopyright} 2022 Halliday, Long, Baum, Thomas, Shelley, Oliver, Gupta, Francis, Williamson, Di Bartolo, Randell, Ben-Khoud, Kelland, Mortimer, Ball, Plumptre, Chandler, Obst, Secchi, Piemonti, Lampasona, Smith, Gregorova, Knezevic, Metz, Barr, Morales-Aza, Oliver, Collingwood, Hitchings, Ring, Wooldridge, Rivino, Timpson, McKernon, Muir, Hamilton, Arnold, Woolfson, Goenka, Davidson, Toye, Berger, Bailey, Gillespie, Williams and Finn.",

year = "2022",

month = nov,

day = "9",

doi = "10.3389/fimmu.2022.968317",

language = "English",

volume = "13",

journal = "Frontiers in Immunology",

issn = "1664-3224",

publisher = "Frontiers Media S.A.",

}

Halliday, A , Long, AE, Baum, HE, Thomas, A, Shelley, KL, Oliver, EH, Gupta, K, Francis, O, Kavanagh Williamson, M, Di Bartolo, ND, Randell, MJ, Ben Khoud, Y, Kelland, I, Mortimer, GLM, Ball, OE, Plumptre, C, Chandler, MA, Obst, U, Secchi, M, Piemonti, L, Lampasona, V, Smith, J, Gregorova, M , Knezevic, L , Metz, J , Barr, RS , Morales-Aza, MB, Oliver, JL, Collingwood, L, Hitchings, BE, Ring, SM , Wooldridge, L , Rivino, L , Timpson, NJ, McKernon, J, Muir, P, Hamilton, FW , Arnold, DT , Woolfson, DN , Goenka, A , Davidson, AD , Toye, AM , Berger, I , Bailey, M , Gillespie, KM, Williams, AJK & Finn, A 2022, 'Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2', Frontiers in Immunology, vol. 13, 968317. https://doi.org/10.3389/fimmu.2022.968317

TY - JOUR

T1 - Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2

AU - Halliday, Alice

AU - Long, Anna E

AU - Baum, Holly E

AU - Thomas, Amy

AU - Shelley, Kathryn L

AU - Oliver, Elizabeth H

AU - Gupta, Kapil

AU - Francis, Ore

AU - Kavanagh Williamson, Maia

AU - Di Bartolo, Natalie D

AU - Randell, Matthew J

AU - Ben Khoud, Yassin

AU - Kelland, Ilana

AU - Mortimer, Georgina L M

AU - Ball, Olivia E

AU - Plumptre, Charlie

AU - Chandler, M A

AU - Obst, Ulrike

AU - Secchi, Massimiliano

AU - Piemonti, Lorenzo

AU - Lampasona, Vito

AU - Smith, Joyce

AU - Gregorova, Michaela

AU - Knezevic, Lea

AU - Metz, Jane

AU - Barr, Rachael S

AU - Morales-Aza, M B

AU - Oliver, Jennifer L

AU - Collingwood, Lucy

AU - Hitchings, Benjamin E

AU - Ring, Susan M

AU - Wooldridge, Linda

AU - Rivino, Laura

AU - Timpson, Nicholas John

AU - McKernon, Jorgen

AU - Muir, Peter

AU - Hamilton, Fergus W

AU - Arnold, David T

AU - Woolfson, Dek N

AU - Goenka, Anu

AU - Davidson, Andrew D

AU - Toye, Ash M

AU - Berger, Imre

AU - Bailey, Mick

AU - Gillespie, Kathleen M

AU - Williams, Alistair J K

AU - Finn, Adam

N1 - Funding Information: This work was supported by multiple grants to AH, AL, OF, AMT, and IB awarded by the Elizabeth Blackwell Institute, and funded in part by the Wellcome Trust [Grant number 204813/Z/16/Z] with additional support from Bristol Alumni and Friends. AL is funded by a Diabetes UK/JDRF RD Lawrence Fellowship (18/0005778 and 3-APF-2018-591-A-N). The DISCOVER study was supported by donations to Southmead Hospital Charity (Registered Charity Number: 1055900). The LOGIC study was funded by The Grand Appeal (The Official Bristol Children’s Hospital Charity; a registered charity in England and Wales (1043603)) through a grant awarded to AF and AG. ACT is supported by the Wellcome Trust (217509/Z/19/Z) and UKRI through the JUNIPER consortium MR/V038613/1 and CoMMinS study MR/V028545/1. The UK Medical Research Council and Wellcome (Grant ref: 217065/Z/19/Z) and the University of Bristol provide core support for ALSPAC. NT is a Wellcome Trust Investigator (202802/Z/16/Z), is the PI of the Avon Longitudinal Study of Parents and Children (MRC & WT 217065/Z/19/Z), is supported by the University of Bristol NIHR Biomedical Research Centre (BRC-1215-2001), the MRC Integrative Epidemiology Unit (MC_UU_00011/1) and works within the CRUK Integrative Cancer Epidemiology Programme (C18281/A29019). LIPS assay development was supported by a joint grant from Diabetes UK/JDRF (20/0006217) to KMG. IB is supported by the Wellcome Trust (106115/Z/14/Z, 221708/Z/20/Z), the ERC (contr. nrs. 834631, 963992) and the EPSRC Impact Acceleration Account EP/R511663/1. We also acknowledge funding from BBSRC/EPSRC Synthetic Biology Research Centre (BB/L01386X/1, to DNW, NB and AMT), NHS Blood and Transplant (WP15-05, to NB and AMT), and the NIHR Blood and Transplant Research Unit in Red Cell Products (IS-BTU-1214-10032, to NB and AMT). This publication is the work of the authors and AH et al. will serve as guarantors for the contents of this paper. Funding Information: AF is a member of the Joint Committee on Vaccination and Immunisation, the UK National Immunisation Technical Advisory Group and is chair of the WHO European Regional Technical Advisory Group of Experts (ETAGE) on immunization and ex officio a member of the WHO SAGE working group on COVID vaccines. He is investigator COVID-19 vaccine on studies and trials funded by Pfizer, Sanofi, Valneva, the Gates Foundation and the UK government. This manuscript presents independent research funded in part by the National Institute for Health Research (NIHR). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health and Social Care. Publisher Copyright: Copyright © 2022 Halliday, Long, Baum, Thomas, Shelley, Oliver, Gupta, Francis, Williamson, Di Bartolo, Randell, Ben-Khoud, Kelland, Mortimer, Ball, Plumptre, Chandler, Obst, Secchi, Piemonti, Lampasona, Smith, Gregorova, Knezevic, Metz, Barr, Morales-Aza, Oliver, Collingwood, Hitchings, Ring, Wooldridge, Rivino, Timpson, McKernon, Muir, Hamilton, Arnold, Woolfson, Goenka, Davidson, Toye, Berger, Bailey, Gillespie, Williams and Finn.

PY - 2022/11/9

Y1 - 2022/11/9

N2 - Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97% specificity and >92% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.

AB - Low-volume antibody assays can be used to track SARS-CoV-2 infection rates in settings where active testing for virus is limited and remote sampling is optimal. We developed 12 ELISAs detecting total or antibody isotypes to SARS-CoV-2 nucleocapsid, spike protein or its receptor binding domain (RBD), 3 anti-RBD isotype specific luciferase immunoprecipitation system (LIPS) assays and a novel Spike-RBD bridging LIPS total-antibody assay. We utilized pre-pandemic (n=984) and confirmed/suspected recent COVID-19 sera taken pre-vaccination rollout in 2020 (n=269). Assays measuring total antibody discriminated best between pre-pandemic and COVID-19 sera and were selected for diagnostic evaluation. In the blind evaluation, two of these assays (Spike Pan ELISA and Spike-RBD Bridging LIPS assay) demonstrated >97% specificity and >92% sensitivity for samples from COVID-19 patients taken >21 days post symptom onset or PCR test. These assays offered better sensitivity for the detection of COVID-19 cases than a commercial assay which requires 100-fold larger serum volumes. This study demonstrates that low-volume in-house antibody assays can provide good diagnostic performance, and highlights the importance of using well-characterized samples and controls for all stages of assay development and evaluation. These cost-effective assays may be particularly useful for seroprevalence studies in low and middle-income countries.

U2 - 10.3389/fimmu.2022.968317

DO - 10.3389/fimmu.2022.968317

M3 - Article (Academic Journal)

C2 - 36439154

SN - 1664-3224

VL - 13

JO - Frontiers in Immunology

JF - Frontiers in Immunology

M1 - 968317

ER -

Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2

Abstract

Bibliographical note

Structured keywords

UN SDGs

Access to Document

Handle.net

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Projects

Elizabeth Blackwell Institute Early Career Fellowship

Datasets

Development and evaluation of low-volume tests to detect and characterize antibodies to SARS-CoV-2

Cite this