A point-of-care lateral flow assay for neutralising antibodies against SARS-CoV-2

Thomas S Fulford; Huy Van; Nicholas A Gherardin; Shuning Zheng; Marcin Ciula; Heidi E Drummer; Samuel Redmond; Hyon-Xhi Tan; Irene Boo; Rob J Center; Fan Li; Samantha L Grimley; Bruce D Wines; Thi H O Nguyen; Francesca L Mordant; Paula Ellenberg; Louise C Rowntree; Lukasz Kedzierski; Allen C Cheng; Denise L Doolan; Gail Matthews; Katherine Bond; P Mark Hogarth; Zoe McQuilten; Kanta Subbarao; Katherine Kedzierska; Jennifer A Juno; Adam K Wheatley; Stephen J Kent; Deborah A Williamson; Damian F J Purcell; David A Anderson; Dale I Godfrey

doi:10.1016/j.ebiom.2021.103729

A point-of-care lateral flow assay for neutralising antibodies against SARS-CoV-2

EBioMedicine. 2021 Dec:74:103729. doi: 10.1016/j.ebiom.2021.103729. Epub 2021 Dec 3.

Authors

Thomas S Fulford¹, Huy Van², Nicholas A Gherardin³, Shuning Zheng², Marcin Ciula¹, Heidi E Drummer⁴, Samuel Redmond¹, Hyon-Xhi Tan¹, Irene Boo², Rob J Center⁵, Fan Li², Samantha L Grimley¹, Bruce D Wines⁶, Thi H O Nguyen¹, Francesca L Mordant¹, Paula Ellenberg¹, Louise C Rowntree¹, Lukasz Kedzierski¹, Allen C Cheng⁷, Denise L Doolan⁸, Gail Matthews⁹, Katherine Bond¹⁰, P Mark Hogarth⁶, Zoe McQuilten⁷, Kanta Subbarao¹¹, Katherine Kedzierska¹, Jennifer A Juno¹, Adam K Wheatley¹², Stephen J Kent¹³, Deborah A Williamson¹⁰, Damian F J Purcell¹, David A Anderson¹⁴, Dale I Godfrey¹⁵

Affiliations

¹ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia.
² Burnet Institute, Melbourne, Victoria, Australia.
³ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Melbourne, Victoria, Australia.
⁴ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Burnet Institute, Melbourne, Victoria, Australia; Department of Microbiology, Monash University, Australia.
⁵ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Burnet Institute, Melbourne, Victoria, Australia.
⁶ Immune therapies Laboratory, Burnet Institute, Melbourne, VIC, Australia,; Department of Immunology and Pathology, Central Clinical School, Monash University, Melbourne, VIC, Australia; Department of Clinical Pathology, The University of Melbourne, Parkville, VIC, Australia.
⁷ Department of Epidemiology and Preventive Medicine, Monash University, Melbourne, Victoria, Australia.
⁸ Australian Institute of Tropical Health & Medicine, James Cook University, Cairns, Queensland, Australia.
⁹ Kirby Institute, University of NSW, Sydney, NSW, Australia.
¹⁰ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Department of Microbiology, Royal Melbourne Hospital, Melbourne, Australia.
¹¹ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; WHO Collaborating Centre for Reference and Research on Influenza at the Peter Doherty Institute for Infection and Immunity.
¹² Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Melbourne Sexual Health Centre and Department of Infectious Diseases, Alfred Hospital and Central Clinical School, Monash University, Melbourne, Victoria, Australia.
¹³ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Melbourne Sexual Health Centre and Department of Infectious Diseases, Alfred Hospital and Central Clinical School, Monash University, Melbourne, Victoria, Australia; Australian Research Council Centre for Excellence in Convergent Bio-Nano Science and Technology, University of Melbourne, Melbourne, Victoria, Australia.
¹⁴ Burnet Institute, Melbourne, Victoria, Australia. Electronic address: david.anderson@burnet.edu.au.
¹⁵ Department of Microbiology and Immunology, University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Victoria, Australia; Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Melbourne, Victoria, Australia. Electronic address: godfrey@unimelb.edu.au.

Abstract

Background: As vaccines against SARS-CoV-2 are now being rolled out, a better understanding of immunity to the virus, whether from infection, or passive or active immunisation, and the durability of this protection is required. This will benefit from the ability to measure antibody-based protection to SARS-CoV-2, ideally with rapid turnaround and without the need for laboratory-based testing.

Methods: We have developed a lateral flow POC test that can measure levels of RBD-ACE2 neutralising antibody (NAb) from whole blood, with a result that can be determined by eye or quantitatively on a small instrument. We compared our lateral flow test with the gold-standard microneutralisation assay, using samples from convalescent and vaccinated donors, as well as immunised macaques.

Findings: We show a high correlation between our lateral flow test with conventional neutralisation and that this test is applicable with animal samples. We also show that this assay is readily adaptable to test for protection to newly emerging SARS-CoV-2 variants, including the beta variant which revealed a marked reduction in NAb activity. Lastly, using a cohort of vaccinated humans, we demonstrate that our whole-blood test correlates closely with microneutralisation assay data (specificity 100% and sensitivity 96% at a microneutralisation cutoff of 1:40) and that fingerprick whole blood samples are sufficient for this test.

Interpretation: Taken together, the COVID-19 NAb-test^TM device described here provides a rapid readout of NAb based protection to SARS-CoV-2 at the point of care.

Funding: Support was received from the Victorian Operational Infrastructure Support Program and the Australian Government Department of Health. This work was supported by grants from the Department of Health and Human Services of the Victorian State Government; the ARC (CE140100011, CE140100036), the NHMRC (1113293, 2002317 and 1116530), and Medical Research Future Fund Awards (2005544, 2002073, 2002132). Individual researchers were supported by an NHMRC Emerging Leadership Level 1 Investigator Grants (1194036), NHMRC APPRISE Research Fellowship (1116530), NHMRC Leadership Investigator Grant (1173871), NHMRC Principal Research Fellowship (1137285), NHMRC Investigator Grants (1177174 and 1174555) and NHMRC Senior Principal Research Fellowships (1117766 and 1136322). Grateful support was also received from the A2 Milk Company and the Jack Ma Foundation.

Keywords: Lateral flow assay; Neturalising antibodies; Point of care test; SARS-CoV-2.

MeSH terms

Animals
Antibodies, Neutralizing / blood*
Antibodies, Viral / blood*
Australia
COVID-19 / immunology*
COVID-19 Serological Testing / methods*
COVID-19 Vaccines / immunology
Humans
Macaca / immunology
Neutralization Tests
Point-of-Care Systems*
SARS-CoV-2 / immunology*
Vaccination

Substances

Antibodies, Neutralizing
Antibodies, Viral
COVID-19 Vaccines

Supplementary concepts

SARS-CoV-2 variants