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 C. Thomas, Kathryn L. Shelley, Elizabeth Oliver, Kapil Gupta, Ore Francis, Maia Kavanagh Williamson, Natalie Di Bartolo, Matthew J. Randell, Yassin Ben-Khoud, Ilana Kelland, Georgina Mortimer, Olivia Ball, Charlie Plumptre, Kyla Chandler, Ulrike Obst, Massimiliano Secchi, Lorenzo PiemontiVito Lampasona, Joyce Smith, Michaela Gregorova, Lea Knezevic, Jane Metz, Rachael Barr, Begonia Morales-Aza, Jennifer Oliver, Lucy Collingwood, Benjamin Hitchings, Susan Ring, Linda Wooldridge, Laura Rivino, Nicholas Timpson, Jorgen McKernon, Peter Muir, Fergus Hamilton, David Arnold, Derek N. Woolfson, Anu Goenka, Andrew D. Davidson, Ashley M. Toye, Imre Berger, Mick Bailey, Kathleen M. Gillespie, Alistair J.K. Williams, Adam Finn

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)


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 languageEnglish
Article number968317
JournalFrontiers in Immunology
Publication statusPublished - 9 Nov 2022
Externally publishedYes

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.


  • COVID-19
  • SARS-CoV-2
  • antibody
  • diagnostic
  • evaluation
  • immunity
  • luciferase immunoprecipitation system (LIPS)


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