Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics

Summary: SARS-CoV-2 continues to evolve, and its emerging variants might escape the immune responses generated by existing vaccines and therapeutic mAbs. Accordingly, rapid analysis of their possible neutralization phenotype is essential and can be facilitated by reverse genetics (RGs) systems to re...

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Main Authors: Madeeha Afzal, Diana Melnyk, Thomas Courty, Lisa Schimanski, Michelle Hill, Stuart Neil, Tiong Kit Tan, William S. James
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:iScience
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589004225007126
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author Madeeha Afzal
Diana Melnyk
Thomas Courty
Lisa Schimanski
Michelle Hill
Stuart Neil
Tiong Kit Tan
William S. James
author_facet Madeeha Afzal
Diana Melnyk
Thomas Courty
Lisa Schimanski
Michelle Hill
Stuart Neil
Tiong Kit Tan
William S. James
author_sort Madeeha Afzal
collection DOAJ
description Summary: SARS-CoV-2 continues to evolve, and its emerging variants might escape the immune responses generated by existing vaccines and therapeutic mAbs. Accordingly, rapid analysis of their possible neutralization phenotype is essential and can be facilitated by reverse genetics (RGs) systems to regenerate viruses with variant-specific substitutions. Here, we efficiently generate a panel of recent variants of SARS-CoV-2 (Omicron XBB.1.16, EG.5.1, BA.2.86, and JN.1) using a substantially optimized circular polymerase extension reaction (CPER) RGs system. Neutralization potency was analyzed for mAbs targeting different regions of spike protein. mAbs P4-J15, C68.61, S2X259, and IY-2A IgG were able to neutralize all recent viruses. However, S309, which was previously used to treat infection and targets the outer face of RBD, showed ∼75-fold reduction in potency versus JN.1. Moreover, C68.59, which targets the SD1 region of the CTD, was unable to neutralize either BA.2.86 or JN1, which share the E554K substitution in SD1. CPER RGs system and microneutralization assays can be adopted as effective tools to evaluate the efficacy of therapeutic mAbs against emerging variants in a time-responsive manner.
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spelling doaj-art-904f2cfa81aa43ba9587c3eeea0fa31c2025-08-20T03:53:47ZengElsevieriScience2589-00422025-06-0128611245110.1016/j.isci.2025.112451Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse geneticsMadeeha Afzal0Diana Melnyk1Thomas Courty2Lisa Schimanski3Michelle Hill4Stuart Neil5Tiong Kit Tan6William S. James7James & Lillian Martin Centre, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE Oxford, UK; Corresponding authorMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, OX3 9DS Oxford, UKDepartment of Infectious Diseases, School of Immunology & Microbial Sciences, King’s College London, SE1 9RT London, UKMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, OX3 9DS Oxford, UKJames & Lillian Martin Centre, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE Oxford, UKDepartment of Infectious Diseases, School of Immunology & Microbial Sciences, King’s College London, SE1 9RT London, UKMRC Translational Immune Discovery Unit, MRC Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, OX3 9DS Oxford, UK; Chinese Academy of Medical Sciences Oxford Institute, Nuffield Department of Medicine, University of Oxford, OX3 7BN Oxford, UKJames & Lillian Martin Centre, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE Oxford, UKSummary: SARS-CoV-2 continues to evolve, and its emerging variants might escape the immune responses generated by existing vaccines and therapeutic mAbs. Accordingly, rapid analysis of their possible neutralization phenotype is essential and can be facilitated by reverse genetics (RGs) systems to regenerate viruses with variant-specific substitutions. Here, we efficiently generate a panel of recent variants of SARS-CoV-2 (Omicron XBB.1.16, EG.5.1, BA.2.86, and JN.1) using a substantially optimized circular polymerase extension reaction (CPER) RGs system. Neutralization potency was analyzed for mAbs targeting different regions of spike protein. mAbs P4-J15, C68.61, S2X259, and IY-2A IgG were able to neutralize all recent viruses. However, S309, which was previously used to treat infection and targets the outer face of RBD, showed ∼75-fold reduction in potency versus JN.1. Moreover, C68.59, which targets the SD1 region of the CTD, was unable to neutralize either BA.2.86 or JN1, which share the E554K substitution in SD1. CPER RGs system and microneutralization assays can be adopted as effective tools to evaluate the efficacy of therapeutic mAbs against emerging variants in a time-responsive manner.http://www.sciencedirect.com/science/article/pii/S2589004225007126Biological sciencesGeneticsImmunologyImmune system evolutionImmunity
spellingShingle Madeeha Afzal
Diana Melnyk
Thomas Courty
Lisa Schimanski
Michelle Hill
Stuart Neil
Tiong Kit Tan
William S. James
Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics
iScience
Biological sciences
Genetics
Immunology
Immune system evolution
Immunity
title Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics
title_full Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics
title_fullStr Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics
title_full_unstemmed Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics
title_short Determination of resilience of a panel of broadly neutralizing mAbs to emerging variants of SARS-CoV-2 generated using reverse genetics
title_sort determination of resilience of a panel of broadly neutralizing mabs to emerging variants of sars cov 2 generated using reverse genetics
topic Biological sciences
Genetics
Immunology
Immune system evolution
Immunity
url http://www.sciencedirect.com/science/article/pii/S2589004225007126
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