Detection and quantification of antibody to SARS CoV 2 receptor binding domain provides enhanced sensitivity, specificity and utility

Carolina Rosadas; Maryam Khan; Eleanor Parker; Federica Marchesin; Ksenia Katsanovskaja; Macià Sureda-Vives; Natalia Fernandez; Paul Randell; Ruth Harvey; Alice Lilley; Benjamin H L Harris; Mohamed Zuhair; Michael Fertleman; Samreen Ijaz; Steve Dicks; Charlotte-Eve Short; Rachael Quinlan; Graham P Taylor; Kai Hu; Paul McKay; Annachiara Rosa; Chloe Roustan; Mark Zuckerman; Kate El Bouzidi; Graham Cooke; Barnaby Flower; Maya Moshe; Paul Elliott; Alexandra J Spencer; Teresa Lambe; Sarah C Gilbert; Hugh Kingston; J Kenneth Baillie; Peter J M Openshaw; Malcolm G Semple; Peter Cherepanov; Myra O McClure; Richard S Tedder; ISARIC4C Investigators

doi:10.1016/j.jviromet.2022.114475

Detection and quantification of antibody to SARS CoV 2 receptor binding domain provides enhanced sensitivity, specificity and utility

J Virol Methods. 2022 Apr:302:114475. doi: 10.1016/j.jviromet.2022.114475. Epub 2022 Jan 22.

Authors

Carolina Rosadas¹, Maryam Khan¹, Eleanor Parker¹, Federica Marchesin¹, Ksenia Katsanovskaja¹, Macià Sureda-Vives¹, Natalia Fernandez¹, Paul Randell², Ruth Harvey³, Alice Lilley³, Benjamin H L Harris⁴, Mohamed Zuhair⁵, Michael Fertleman⁵, Samreen Ijaz⁶, Steve Dicks⁷, Charlotte-Eve Short¹, Rachael Quinlan¹, Graham P Taylor¹, Kai Hu¹, Paul McKay¹, Annachiara Rosa⁸, Chloe Roustan⁹, Mark Zuckerman¹⁰, Kate El Bouzidi¹⁰, Graham Cooke¹, Barnaby Flower¹, Maya Moshe¹, Paul Elliott¹¹, Alexandra J Spencer¹², Teresa Lambe¹², Sarah C Gilbert¹², Hugh Kingston², J Kenneth Baillie¹³, Peter J M Openshaw¹⁴, Malcolm G Semple¹⁵, Peter Cherepanov¹⁶, Myra O McClure¹, Richard S Tedder¹⁷; ISARIC4C Investigators

Affiliations

¹ Department of Infectious Disease, Imperial College London, St Mary's Campus, London, W2 1PG, UK.
² Department of Infection and Immunity, Imperial College Healthcare NHS Trust, Charing Cross Hospital, London, W6 8RF, UK.
³ Worldwide Influenza Centre, Francis Crick Institute, London, NW1 1AT, UK.
⁴ The Wellington Hospital, Circus Road, St John's Wood, London, NW8 6PD, UK; Computational Biology and Integrative Genomics, Department of Oncology, University of Oxford, Oxford, OX3 7DQ, UK.
⁵ The Wellington Hospital, Circus Road, St John's Wood, London, NW8 6PD, UK.
⁶ Blood Borne Virus Unit, National Infection Service, Colindale Public Health England, London, NW9 5EQ, UK.
⁷ Blood Borne Virus Unit, National Infection Service, Colindale Public Health England, London, NW9 5EQ, UK; Transfusion Microbiology, NHS Blood and Transplant, Lingard Avenue, London, NW9 5BG, UK.
⁸ Chromatin Structure and Mobile DNA Laboratory, The Francis Crick Institute, London, NW1 1AT, UK; Crick COVID19 Consortium, Francis Crick Institute, London, NW1 1AT, UK.
⁹ Structural Biology Science Technology Platform, Francis Crick Institute, London, NW1 1AT, UK; Crick COVID19 Consortium, Francis Crick Institute, London, NW1 1AT, UK.
¹⁰ Department of Virology, King's College Hospital, London, SE5 9RS, UK.
¹¹ Department of Epidemiology and Biostatistics, School of Public Health, Imperial College London, St Mary's Campus, London, W2 1PG, UK.
¹² Jenner Institute, University of Oxford, ORCRB, Oxford, OX3 7DQ, UK.
¹³ Roslin Institute, University of Edinburgh, Midlothian, EH25 9RG, UK.
¹⁴ National Heart and Lung Institute, Imperial College London, Chelsea, London, SW3 6LY, UK.
¹⁵ NIHR Health Protection Research Unit in Emerging and Zoonotic Infections, Institute of Infection, Veterinary and Ecological Sciences, Faculty of Health and Life Sciences, University of Liverpool, Liverpool, L69 7BE, UK.
¹⁶ Department of Infectious Disease, Imperial College London, St Mary's Campus, London, W2 1PG, UK; Chromatin Structure and Mobile DNA Laboratory, The Francis Crick Institute, London, NW1 1AT, UK; Crick COVID19 Consortium, Francis Crick Institute, London, NW1 1AT, UK.
¹⁷ Department of Infectious Disease, Imperial College London, St Mary's Campus, London, W2 1PG, UK. Electronic address: r.tedder@imperial.ac.uk.

Abstract

Accurate and sensitive detection of antibody to SARS-CoV-2 remains an essential component of the pandemic response. Measuring antibody that predicts neutralising activity and the vaccine response is an absolute requirement for laboratory-based confirmatory and reference activity. The viral receptor binding domain (RBD) constitutes the prime target antigen for neutralising antibody. A double antigen binding assay (DABA), providing the most sensitive format has been exploited in a novel hybrid manner employing a solid-phase S1 preferentially presenting RBD, coupled with a labelled RBD conjugate, used in a two-step sequential assay for detection and measurement of antibody to RBD (anti-RBD). This class and species neutral assay showed a specificity of 100 % on 825 pre COVID-19 samples and a potential sensitivity of 99.6 % on 276 recovery samples, predicting quantitatively the presence of neutralising antibody determined by pseudo-type neutralization and by plaque reduction. Anti-RBD is also measurable in ferrets immunised with ChadOx1 nCoV-19 vaccine and in humans immunised with both AstraZeneca and Pfizer vaccines. This assay detects anti-RBD at presentation with illness, demonstrates its elevation with disease severity, its sequel to asymptomatic infection and its persistence after the loss of antibody to the nucleoprotein (anti-NP). It also provides serological confirmation of prior infection and offers a secure measure for seroprevalence and studies of vaccine immunisation in human and animal populations. The hybrid DABA also displays the attributes necessary for the detection and quantification of anti-RBD to be used in clinical practice. An absence of detectable anti-RBD by this assay predicates the need for passive immune prophylaxis in at-risk patients.

Keywords: Antibodies; ELISA; Receptor binding domain; Sars-CoV-2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Neutralizing / isolation & purification
Antibodies, Viral / isolation & purification*
COVID-19* / diagnosis
ChAdOx1 nCoV-19
Ferrets
Humans
RNA, Viral
SARS-CoV-2*
Seroepidemiologic Studies
Spike Glycoprotein, Coronavirus / immunology*

Substances

Antibodies, Neutralizing
Antibodies, Viral
RNA, Viral
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2
ChAdOx1 nCoV-19

Abstract

Publication types

MeSH terms

Substances

Grants and funding