IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy

Despite the unique advantages of IgG3 over other IgG subclasses, such as mediating enhanced effector functions and increased flexibility in antigen binding due to a long hinge region, the therapeutic potential of IgG3 remains largely unexplored. This may be attributed to difficulties in recombinant...

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Main Authors: Somanath Kallolimath, Lin Sun, Roman Palt, Esther Föderl-Höbenreich, Antonia Hermle, Leonie Voss, Marina Kleim, Falk Nimmerjahn, Johannes S. Gach, Lauren Hitchcock, Qiang Chen, Stanislav Melnik, Florian Eminger, Anja Lux, Herta Steinkellner
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-10-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2024.1490515/full
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author Somanath Kallolimath
Lin Sun
Roman Palt
Esther Föderl-Höbenreich
Antonia Hermle
Leonie Voss
Marina Kleim
Falk Nimmerjahn
Falk Nimmerjahn
Johannes S. Gach
Lauren Hitchcock
Qiang Chen
Stanislav Melnik
Florian Eminger
Anja Lux
Anja Lux
Herta Steinkellner
author_facet Somanath Kallolimath
Lin Sun
Roman Palt
Esther Föderl-Höbenreich
Antonia Hermle
Leonie Voss
Marina Kleim
Falk Nimmerjahn
Falk Nimmerjahn
Johannes S. Gach
Lauren Hitchcock
Qiang Chen
Stanislav Melnik
Florian Eminger
Anja Lux
Anja Lux
Herta Steinkellner
author_sort Somanath Kallolimath
collection DOAJ
description Despite the unique advantages of IgG3 over other IgG subclasses, such as mediating enhanced effector functions and increased flexibility in antigen binding due to a long hinge region, the therapeutic potential of IgG3 remains largely unexplored. This may be attributed to difficulties in recombinant expression and the reduced plasma half-life of most IgG3 allotypes. Here, we report plant expression of two SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) that exhibit high (P5C3) and low (H4) antigen binding. P5C3 and H4-IgG1 mAbs were subclass-switched to IgG3 formats, designed for efficient production and increased PK values, carrying three allotypic variations, referred to as -WT, -H, and -KVH. A total of eight mAbs were produced in glycoengineered plants that synthesize fucose-free complex N-glycans with great homogeneity. Antigen, IgG-FcγR immune complex and complement binding studies demonstrated similar activities of all mAbs. In accordance, P5C3 Abs showed minor alterations in SARS-CoV-2 neutralization (NT) and antibody-dependent cell-mediated virus inhibition (ADCVI). Clear functional differences were observed between H4 variants with superior ADCVI and NT potencies of H4 IgG3 H. Our comparative study demonstrates the production of an IgG3 variant carrying an Fc domain with equivalent or enhanced functions compared to IgG3-WT, but with the stability and PK values ​​of IgG1. Our data also demonstrate that both allotypic variability and antibody specificity are important for fine-tuning of activities, an important information for the development of future therapeutics.
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spelling doaj-art-a923dba30332487ea742f0b863b2b29e2025-08-20T01:54:16ZengFrontiers Media S.A.Frontiers in Immunology1664-32242024-10-011510.3389/fimmu.2024.14905151490515IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacySomanath Kallolimath0Lin Sun1Roman Palt2Esther Föderl-Höbenreich3Antonia Hermle4Leonie Voss5Marina Kleim6Falk Nimmerjahn7Falk Nimmerjahn8Johannes S. Gach9Lauren Hitchcock10Qiang Chen11Stanislav Melnik12Florian Eminger13Anja Lux14Anja Lux15Herta Steinkellner16Institute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, BOKU University, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, BOKU University, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, BOKU University, Vienna, AustriaDiagnostic and Research Institute of Pathology, Medical University of Graz, Graz, AustriaDivision of Genetics, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyDivision of Genetics, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyDivision of Genetics, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyDivision of Genetics, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyFAU Profile Centre Immunomedicine, Erlangen, GermanyDivision of Infectious Diseases, University of California, Irvine, Irvine, CA, United StatesDivision of Infectious Diseases, University of California, Irvine, Irvine, CA, United StatesThe Bio design Institute and School of Life Sciences, Arizona State University, Tempe, AZ, United StatesInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, BOKU University, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, BOKU University, Vienna, AustriaDivision of Genetics, Department of Biology, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, GermanyFAU Profile Centre Immunomedicine, Erlangen, GermanyInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, BOKU University, Vienna, AustriaDespite the unique advantages of IgG3 over other IgG subclasses, such as mediating enhanced effector functions and increased flexibility in antigen binding due to a long hinge region, the therapeutic potential of IgG3 remains largely unexplored. This may be attributed to difficulties in recombinant expression and the reduced plasma half-life of most IgG3 allotypes. Here, we report plant expression of two SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) that exhibit high (P5C3) and low (H4) antigen binding. P5C3 and H4-IgG1 mAbs were subclass-switched to IgG3 formats, designed for efficient production and increased PK values, carrying three allotypic variations, referred to as -WT, -H, and -KVH. A total of eight mAbs were produced in glycoengineered plants that synthesize fucose-free complex N-glycans with great homogeneity. Antigen, IgG-FcγR immune complex and complement binding studies demonstrated similar activities of all mAbs. In accordance, P5C3 Abs showed minor alterations in SARS-CoV-2 neutralization (NT) and antibody-dependent cell-mediated virus inhibition (ADCVI). Clear functional differences were observed between H4 variants with superior ADCVI and NT potencies of H4 IgG3 H. Our comparative study demonstrates the production of an IgG3 variant carrying an Fc domain with equivalent or enhanced functions compared to IgG3-WT, but with the stability and PK values ​​of IgG1. Our data also demonstrate that both allotypic variability and antibody specificity are important for fine-tuning of activities, an important information for the development of future therapeutics.https://www.frontiersin.org/articles/10.3389/fimmu.2024.1490515/fullIgG3 allotypesplant expressionSARS-CoV-2 antibodiesfunctional activitiesantibody engineering
spellingShingle Somanath Kallolimath
Lin Sun
Roman Palt
Esther Föderl-Höbenreich
Antonia Hermle
Leonie Voss
Marina Kleim
Falk Nimmerjahn
Falk Nimmerjahn
Johannes S. Gach
Lauren Hitchcock
Qiang Chen
Stanislav Melnik
Florian Eminger
Anja Lux
Anja Lux
Herta Steinkellner
IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy
Frontiers in Immunology
IgG3 allotypes
plant expression
SARS-CoV-2 antibodies
functional activities
antibody engineering
title IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy
title_full IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy
title_fullStr IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy
title_full_unstemmed IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy
title_short IgG1 versus IgG3: influence of antibody-specificity and allotypic variance on virus neutralization efficacy
title_sort igg1 versus igg3 influence of antibody specificity and allotypic variance on virus neutralization efficacy
topic IgG3 allotypes
plant expression
SARS-CoV-2 antibodies
functional activities
antibody engineering
url https://www.frontiersin.org/articles/10.3389/fimmu.2024.1490515/full
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