Robust correlations across six SARS-CoV-2 serology assays detecting distinct antibody features

Louise C Rowntree; Brendon Y Chua; Suellen Nicholson; Marios Koutsakos; Luca Hensen; Celia Douros; Kevin Selva; Francesca L Mordant; Chinn Yi Wong; Jennifer R Habel; Wuji Zhang; Xiaoxiao Jia; Lily Allen; Denise L Doolan; David C Jackson; Adam K Wheatley; Stephen J Kent; Fatima Amanat; Florian Krammer; Kanta Subbarao; Allen C Cheng; Amy W Chung; Mike Catton; Thi Ho Nguyen; Carolien E van de Sandt; Katherine Kedzierska

doi:10.1002/cti2.1258

Robust correlations across six SARS-CoV-2 serology assays detecting distinct antibody features

Clin Transl Immunology. 2021 Feb 28;10(3):e1258. doi: 10.1002/cti2.1258. eCollection 2021.

Authors

Louise C Rowntree¹, Brendon Y Chua^{1

2}, Suellen Nicholson³, Marios Koutsakos¹, Luca Hensen¹, Celia Douros³, Kevin Selva¹, Francesca L Mordant¹, Chinn Yi Wong¹, Jennifer R Habel¹, Wuji Zhang¹, Xiaoxiao Jia¹, Lily Allen¹, Denise L Doolan⁴, David C Jackson^{1

2}, Adam K Wheatley^{1

5}, Stephen J Kent^{1

5

6}, Fatima Amanat^{7

8}, Florian Krammer⁷, Kanta Subbarao^{1

9}, Allen C Cheng^{10

11}, Amy W Chung¹, Mike Catton³, Thi Ho Nguyen¹, Carolien E van de Sandt^{1

12}, Katherine Kedzierska^{1

2}

Affiliations

¹ Department of Microbiology and Immunology University of Melbourne, at the Peter Doherty Institute for Infection and Immunity Melbourne VIC Australia.
² Global Station for Zoonosis Control Global Institution for Collaborative Research and Education (GI-CoRE) Hokkaido University Sapporo Hokkaido Japan.
³ Victorian Infectious Diseases Reference Laboratory The Royal Melbourne Hospital at The Peter Doherty Institute for Infection and Immunity Melbourne VIC Australia.
⁴ Centre for Molecular Therapeutics Australian Institute of Tropical Health & Medicine James Cook University Cairns QLD Australia.
⁵ ARC Centre of Excellence in Convergent Bio-Nano Science and Technology University of Melbourne Melbourne VIC Australia.
⁶ Infectious Diseases Department Melbourne Sexual Health Centre Alfred Health Central Clinical School Monash University Melbourne VIC Australia.
⁷ Department of Microbiology Icahn School of Medicine at Mount Sinai New York NY USA.
⁸ Graduate School of Biomedical Sciences Icahn School of Medicine at Mount Sinai New York NY USA.
⁹ World Health Organisation (WHO) Collaborating Centre for Reference and Research on Influenza, at The Peter Doherty Institute for Infection and Immunity Melbourne VIC Australia.
¹⁰ School of Public Health and Preventive Medicine Monash University Melbourne VIC Australia.
¹¹ Infection Prevention and Healthcare Epidemiology Unit Alfred Health Melbourne VIC Australia.
¹² Department of Hematopoiesis Sanquin Research and Landsteiner Laboratory Amsterdam UMC University of Amsterdam Amsterdam The Netherlands.

Abstract

Objectives: As the world transitions into a new era of the COVID-19 pandemic in which vaccines become available, there is an increasing demand for rapid reliable serological testing to identify individuals with levels of immunity considered protective by infection or vaccination.

Methods: We used 34 SARS-CoV-2 samples to perform a rapid surrogate virus neutralisation test (sVNT), applicable to many laboratories as it circumvents the need for biosafety level-3 containment. We correlated results from the sVNT with five additional commonly used SARS-CoV-2 serology techniques: the microneutralisation test (MNT), in-house ELISAs, commercial Euroimmun- and Wantai-based ELISAs (RBD, spike and nucleoprotein; IgG, IgA and IgM), antigen-binding avidity, and high-throughput multiplex analyses to profile isotype, subclass and Fc effector binding potential. We correlated antibody levels with antibody-secreting cell (ASC) and circulatory T follicular helper (cTfh) cell numbers.

Results: Antibody data obtained with commercial ELISAs closely reflected results using in-house ELISAs against RBD and spike. A correlation matrix across ten measured ELISA parameters revealed positive correlations for all factors. The frequency of inhibition by rapid sVNT strongly correlated with spike-specific IgG and IgA titres detected by both commercial and in-house ELISAs, and MNT titres. Multiplex analyses revealed strongest correlations between IgG, IgG1, FcR and C1q specific to spike and RBD. Acute cTfh-type 1 cell numbers correlated with spike and RBD-specific IgG antibodies measured by ELISAs and sVNT.

Conclusion: Our comprehensive analyses provide important insights into SARS-CoV-2 humoral immunity across distinct serology assays and their applicability for specific research and/or diagnostic questions to assess SARS-CoV-2-specific humoral responses.

Keywords: ELISA; SARS‐CoV‐2 antibodies; T follicular helper cells; antibody‐secreting cells; neutralisation assay.