A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies

Neale Harrison; Lauren Richardson; Chiara Pallini; Ines Morano; Elizabeth Jinks; Jamie Cowley; Hujo Chan; Harriet J. Hill; Aekkachai Tuekprakhon; Zhi Li; Cristina Matas de las Heras; Ana Teodosio; Andrea S. Lavado; Robert Moring; Ayesha Ashraf; Timothy R. Dafforn; Dimitris K. Grammatopoulos; John Gordon; Catherine A. Brady; Lawrence S. Young; Nicholas M. Barnes; Zania Stamataki; Omar S. Qureshi

doi:10.3389/fviro.2023.1163385

A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies

Neale Harrison, Lauren Richardson, Chiara Pallini, Ines Morano, Elizabeth Jinks, Jamie Cowley, Hujo Chan, Harriet J. Hill, Aekkachai Tuekprakhon, Zhi Li, Cristina Matas de las Heras, Ana Teodosio, Andrea S. Lavado, Robert Moring, Ayesha Ashraf, Timothy R. Dafforn, Dimitris K. Grammatopoulos, John Gordon, Catherine A. Brady, Lawrence S. YoungNicholas M. Barnes, Zania Stamataki, Omar S. Qureshi^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

Introduction: The engagement of the SARS-CoV-2 spike protein with ACE2 is a critical step for viral entry to human cells, and, therefore, blocking this interaction is a major determinant of the efficacy of monoclonal antibody therapeutics and vaccine elicited serum antibodies. The emergence of SARS-CoV-2 variants has necessitated the development of adaptable assays that can be applied to assess the effectiveness of antibody-based therapeutics.

Methods: Through the testing of a range of recombinant spike proteins, we have developed a cell-based, ACE2/spike protein interaction assay that characterises monoclonal anti-spike protein antibodies and neutralising antibodies in donor serum. The assay uses high-content imaging to quantify cell-bound spike protein fluorescence.

Results: Using spike proteins from the original “Wuhan” SARS-CoV-2 strain and the Delta and Omicron variants, we identified differential blocking activity of three monoclonal antibodies directed against the spike receptor-binding domain. Importantly, biological activity in the spike interaction assay translated to efficacy in a SARS-CoV-2 infection assay.

Discussion: The spike protein interaction assay can be used to monitor anti-spike antibodies against the major known SARS-CoV-2 variants and is readily adaptable for quantification of the impact of antibodies against new and emerging SARS-CoV-2 variants.

Original language	English
Article number	1163385
Number of pages	12
Journal	Frontiers in Virology
Volume	3
DOIs	https://doi.org/10.3389/fviro.2023.1163385
Publication status	Published - 29 Aug 2023

Keywords

antibody discovery
spike protein
antiviral
virology
SARS-CoV-2
variant
ACE2

UN SDGs

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

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Development of a SARS-CoV-2 spike protein binding assay using human epithelial cells for COVID-19 therapy discovery and development
Willcox, B., Dafforn, T., Barnes, N. & Stamataki, Z.
TECHNOLOGY STRATEGY BOARD
1/01/21 → 30/06/22
Project: Other Government Departments

Cite this

Harrison, N., Richardson, L., Pallini, C., Morano, I., Jinks, E., Cowley, J., Chan, H., Hill, H. J., Tuekprakhon, A., Li, Z., Matas de las Heras, C., Teodosio, A., Lavado, A. S., Moring, R., Ashraf, A., Dafforn, T. R., Grammatopoulos, D. K., Gordon, J., Brady, C. A., ... Qureshi, O. S. (2023). A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies. Frontiers in Virology, 3, Article 1163385. https://doi.org/10.3389/fviro.2023.1163385

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title = "A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies",

abstract = "Introduction: The engagement of the SARS-CoV-2 spike protein with ACE2 is a critical step for viral entry to human cells, and, therefore, blocking this interaction is a major determinant of the efficacy of monoclonal antibody therapeutics and vaccine elicited serum antibodies. The emergence of SARS-CoV-2 variants has necessitated the development of adaptable assays that can be applied to assess the effectiveness of antibody-based therapeutics. Methods: Through the testing of a range of recombinant spike proteins, we have developed a cell-based, ACE2/spike protein interaction assay that characterises monoclonal anti-spike protein antibodies and neutralising antibodies in donor serum. The assay uses high-content imaging to quantify cell-bound spike protein fluorescence. Results: Using spike proteins from the original “Wuhan” SARS-CoV-2 strain and the Delta and Omicron variants, we identified differential blocking activity of three monoclonal antibodies directed against the spike receptor-binding domain. Importantly, biological activity in the spike interaction assay translated to efficacy in a SARS-CoV-2 infection assay. Discussion: The spike protein interaction assay can be used to monitor anti-spike antibodies against the major known SARS-CoV-2 variants and is readily adaptable for quantification of the impact of antibodies against new and emerging SARS-CoV-2 variants.",

keywords = "antibody discovery, spike protein, antiviral, virology, SARS-CoV-2, variant, ACE2",

author = "Neale Harrison and Lauren Richardson and Chiara Pallini and Ines Morano and Elizabeth Jinks and Jamie Cowley and Hujo Chan and Hill, {Harriet J.} and Aekkachai Tuekprakhon and Zhi Li and {Matas de las Heras}, Cristina and Ana Teodosio and Lavado, {Andrea S.} and Robert Moring and Ayesha Ashraf and Dafforn, {Timothy R.} and Grammatopoulos, {Dimitris K.} and John Gordon and Brady, {Catherine A.} and Young, {Lawrence S.} and Barnes, {Nicholas M.} and Zania Stamataki and Qureshi, {Omar S.}",

year = "2023",

month = aug,

day = "29",

doi = "10.3389/fviro.2023.1163385",

language = "English",

volume = "3",

journal = "Frontiers in Virology",

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publisher = "Frontiers Media S.A.",

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Harrison, N, Richardson, L, Pallini, C, Morano, I, Jinks, E, Cowley, J, Chan, H, Hill, HJ, Tuekprakhon, A, Li, Z, Matas de las Heras, C, Teodosio, A, Lavado, AS, Moring, R, Ashraf, A, Dafforn, TR, Grammatopoulos, DK, Gordon, J, Brady, CA, Young, LS, Barnes, NM , Stamataki, Z & Qureshi, OS 2023, 'A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies', Frontiers in Virology, vol. 3, 1163385. https://doi.org/10.3389/fviro.2023.1163385

TY - JOUR

T1 - A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies

AU - Harrison, Neale

AU - Richardson, Lauren

AU - Pallini, Chiara

AU - Morano, Ines

AU - Jinks, Elizabeth

AU - Cowley, Jamie

AU - Chan, Hujo

AU - Hill, Harriet J.

AU - Tuekprakhon, Aekkachai

AU - Li, Zhi

AU - Matas de las Heras, Cristina

AU - Teodosio, Ana

AU - Lavado, Andrea S.

AU - Moring, Robert

AU - Ashraf, Ayesha

AU - Dafforn, Timothy R.

AU - Grammatopoulos, Dimitris K.

AU - Gordon, John

AU - Brady, Catherine A.

AU - Young, Lawrence S.

AU - Barnes, Nicholas M.

AU - Stamataki, Zania

AU - Qureshi, Omar S.

PY - 2023/8/29

Y1 - 2023/8/29

N2 - Introduction: The engagement of the SARS-CoV-2 spike protein with ACE2 is a critical step for viral entry to human cells, and, therefore, blocking this interaction is a major determinant of the efficacy of monoclonal antibody therapeutics and vaccine elicited serum antibodies. The emergence of SARS-CoV-2 variants has necessitated the development of adaptable assays that can be applied to assess the effectiveness of antibody-based therapeutics. Methods: Through the testing of a range of recombinant spike proteins, we have developed a cell-based, ACE2/spike protein interaction assay that characterises monoclonal anti-spike protein antibodies and neutralising antibodies in donor serum. The assay uses high-content imaging to quantify cell-bound spike protein fluorescence. Results: Using spike proteins from the original “Wuhan” SARS-CoV-2 strain and the Delta and Omicron variants, we identified differential blocking activity of three monoclonal antibodies directed against the spike receptor-binding domain. Importantly, biological activity in the spike interaction assay translated to efficacy in a SARS-CoV-2 infection assay. Discussion: The spike protein interaction assay can be used to monitor anti-spike antibodies against the major known SARS-CoV-2 variants and is readily adaptable for quantification of the impact of antibodies against new and emerging SARS-CoV-2 variants.

AB - Introduction: The engagement of the SARS-CoV-2 spike protein with ACE2 is a critical step for viral entry to human cells, and, therefore, blocking this interaction is a major determinant of the efficacy of monoclonal antibody therapeutics and vaccine elicited serum antibodies. The emergence of SARS-CoV-2 variants has necessitated the development of adaptable assays that can be applied to assess the effectiveness of antibody-based therapeutics. Methods: Through the testing of a range of recombinant spike proteins, we have developed a cell-based, ACE2/spike protein interaction assay that characterises monoclonal anti-spike protein antibodies and neutralising antibodies in donor serum. The assay uses high-content imaging to quantify cell-bound spike protein fluorescence. Results: Using spike proteins from the original “Wuhan” SARS-CoV-2 strain and the Delta and Omicron variants, we identified differential blocking activity of three monoclonal antibodies directed against the spike receptor-binding domain. Importantly, biological activity in the spike interaction assay translated to efficacy in a SARS-CoV-2 infection assay. Discussion: The spike protein interaction assay can be used to monitor anti-spike antibodies against the major known SARS-CoV-2 variants and is readily adaptable for quantification of the impact of antibodies against new and emerging SARS-CoV-2 variants.

KW - antibody discovery

KW - spike protein

KW - antiviral

KW - virology

KW - SARS-CoV-2

KW - variant

KW - ACE2

U2 - 10.3389/fviro.2023.1163385

DO - 10.3389/fviro.2023.1163385

M3 - Article

SN - 2673-818X

VL - 3

JO - Frontiers in Virology

JF - Frontiers in Virology

M1 - 1163385

ER -

A cell-based, SARS-CoV-2 spike protein interaction assay to inform the neutralising capacity of recombinant and patient sera antibodies

Abstract

Keywords

UN SDGs

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Development of a SARS-CoV-2 spike protein binding assay using human epithelial cells for COVID-19 therapy discovery and development

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