Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus

ABSTRACT Crimean-Congo hemorrhagic fever virus (CCHFV) is an enveloped, negative-sense RNA virus that is spread by ticks across Europe, Africa, and Asia and causes a lethal disease in humans (~30%–40% case fatality). There are currently no approved vaccines or therapeutics. Antibody-based therapeuti...

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Main Authors: Albert Wang, Stephanie R. Monticelli, Ariel S. Wirchnianski, Dafna M. Abelson, Ana I. Kuehne, Russell R. Bakken, Marissa Middlecamp, Michael Weingart, Olivia Vergnolle, Zachary A. Bornholdt, Crystal L. Moyer, Jacob L. Berrigan, Brandyn R. West, J. Maximilian Fels, Larry Zeitlin, Andrew S. Herbert, Kartik Chandran, Chowdhury Raihan Bikash, Jonathan R. Lai
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Language:English
Published: American Society for Microbiology 2025-05-01
Series:mBio
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Online Access:https://journals.asm.org/doi/10.1128/mbio.03202-24
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author Albert Wang
Stephanie R. Monticelli
Ariel S. Wirchnianski
Dafna M. Abelson
Ana I. Kuehne
Russell R. Bakken
Marissa Middlecamp
Michael Weingart
Olivia Vergnolle
Zachary A. Bornholdt
Crystal L. Moyer
Jacob L. Berrigan
Brandyn R. West
J. Maximilian Fels
Larry Zeitlin
Andrew S. Herbert
Kartik Chandran
Chowdhury Raihan Bikash
Jonathan R. Lai
author_facet Albert Wang
Stephanie R. Monticelli
Ariel S. Wirchnianski
Dafna M. Abelson
Ana I. Kuehne
Russell R. Bakken
Marissa Middlecamp
Michael Weingart
Olivia Vergnolle
Zachary A. Bornholdt
Crystal L. Moyer
Jacob L. Berrigan
Brandyn R. West
J. Maximilian Fels
Larry Zeitlin
Andrew S. Herbert
Kartik Chandran
Chowdhury Raihan Bikash
Jonathan R. Lai
author_sort Albert Wang
collection DOAJ
description ABSTRACT Crimean-Congo hemorrhagic fever virus (CCHFV) is an enveloped, negative-sense RNA virus that is spread by ticks across Europe, Africa, and Asia and causes a lethal disease in humans (~30%–40% case fatality). There are currently no approved vaccines or therapeutics. Antibody-based therapeutics targeting the CCHFV surface glycoproteins Gn and Gc, which are responsible for viral attachment and fusion during entry, are a promising therapeutic approach. We previously isolated three broadly neutralizing Gc-targeting human monoclonal antibodies (mAbs) and showed certain cocktails of these mAbs demonstrated synergistic virus neutralization. Furthermore, physical linkage of two of these mAbs into a dual variable domain (DVD) bispecific antibody (bsAb) DVD-121-801 resulted in improved neutralization and therapeutic protection against a lethal CCHFV challenge in mice. However, the molecular requirements for the activity of DVD-121-801, and why it is augmented over monospecific parental mAbs, remain the topic of investigation. Here, we generated a new panel of bsAb variants of DVD-121-801 to explore the spacing and avidity requirements and further optimize its protective efficacy against divergent CCHFV isolates. We evaluated these variants for neutralization, fusion inhibition, and protection with virus-like particles, authentic viruses, and in vivo challenge studies. We found that neutralization potency was relatively unaffected by spacing or identity of variable domains within the bsAb, but that one next-generation design employing longer and more flexible linkers between variable domains (DVD-121-801GS) had a greater breadth of therapeutic protection. Our efforts highlight the importance of antibody avidity and lead to an improved bsAb variant of DVD-121-801 for further therapeutic development.IMPORTANCECrimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus endemic to Europe, Africa, and Asia that causes severe disease in humans (30%–40% case fatality). There are currently no approved vaccines or therapeutics. In prior work, physical linkage of two Gc-specific monoclonal antibodies (mAbs) targeting distinct epitopes into a dual variable domain (DVD) bispecific antibody (bsAb), termed DVD-121-801, resulted in potent neutralization in vitro and therapeutic protection in vivo. Here, a panel of variants of this bsAb was developed and evaluated for neutralization potency, fusion inhibition, and therapeutic efficacy, and our work shows this panel to be effective against multiple isolates of CCHFV. Furthermore, incorporating longer, more flexible linkers between variable domains resulted in a lead candidate with improved activity and therapeutic potential compared to the parental bsAb. Utilizing this panel, we also explored the contribution of antibody avidity in antibody-mediated protection against CCHFV infection.
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spelling doaj-art-f4b5950fdb5f45b2b6dc46e079a797522025-08-20T03:49:32ZengAmerican Society for MicrobiologymBio2150-75112025-05-0116510.1128/mbio.03202-24Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virusAlbert Wang0Stephanie R. Monticelli1Ariel S. Wirchnianski2Dafna M. Abelson3Ana I. Kuehne4Russell R. Bakken5Marissa Middlecamp6Michael Weingart7Olivia Vergnolle8Zachary A. Bornholdt9Crystal L. Moyer10Jacob L. Berrigan11Brandyn R. West12J. Maximilian Fels13Larry Zeitlin14Andrew S. Herbert15Kartik Chandran16Chowdhury Raihan Bikash17Jonathan R. Lai18Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USAVirology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USAMapp Biopharmaceutical, Inc., San Diego, California, USAVirology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, USAVirology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, USAMapp Biopharmaceutical, Inc., San Diego, California, USAMapp Biopharmaceutical, Inc., San Diego, California, USADepartment of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, USAMapp Biopharmaceutical, Inc., San Diego, California, USAMapp Biopharmaceutical, Inc., San Diego, California, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USAMapp Biopharmaceutical, Inc., San Diego, California, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USAMapp Biopharmaceutical, Inc., San Diego, California, USAVirology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, USADepartment of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USADepartment of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, USADepartment of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, USAABSTRACT Crimean-Congo hemorrhagic fever virus (CCHFV) is an enveloped, negative-sense RNA virus that is spread by ticks across Europe, Africa, and Asia and causes a lethal disease in humans (~30%–40% case fatality). There are currently no approved vaccines or therapeutics. Antibody-based therapeutics targeting the CCHFV surface glycoproteins Gn and Gc, which are responsible for viral attachment and fusion during entry, are a promising therapeutic approach. We previously isolated three broadly neutralizing Gc-targeting human monoclonal antibodies (mAbs) and showed certain cocktails of these mAbs demonstrated synergistic virus neutralization. Furthermore, physical linkage of two of these mAbs into a dual variable domain (DVD) bispecific antibody (bsAb) DVD-121-801 resulted in improved neutralization and therapeutic protection against a lethal CCHFV challenge in mice. However, the molecular requirements for the activity of DVD-121-801, and why it is augmented over monospecific parental mAbs, remain the topic of investigation. Here, we generated a new panel of bsAb variants of DVD-121-801 to explore the spacing and avidity requirements and further optimize its protective efficacy against divergent CCHFV isolates. We evaluated these variants for neutralization, fusion inhibition, and protection with virus-like particles, authentic viruses, and in vivo challenge studies. We found that neutralization potency was relatively unaffected by spacing or identity of variable domains within the bsAb, but that one next-generation design employing longer and more flexible linkers between variable domains (DVD-121-801GS) had a greater breadth of therapeutic protection. Our efforts highlight the importance of antibody avidity and lead to an improved bsAb variant of DVD-121-801 for further therapeutic development.IMPORTANCECrimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus endemic to Europe, Africa, and Asia that causes severe disease in humans (30%–40% case fatality). There are currently no approved vaccines or therapeutics. In prior work, physical linkage of two Gc-specific monoclonal antibodies (mAbs) targeting distinct epitopes into a dual variable domain (DVD) bispecific antibody (bsAb), termed DVD-121-801, resulted in potent neutralization in vitro and therapeutic protection in vivo. Here, a panel of variants of this bsAb was developed and evaluated for neutralization potency, fusion inhibition, and therapeutic efficacy, and our work shows this panel to be effective against multiple isolates of CCHFV. Furthermore, incorporating longer, more flexible linkers between variable domains resulted in a lead candidate with improved activity and therapeutic potential compared to the parental bsAb. Utilizing this panel, we also explored the contribution of antibody avidity in antibody-mediated protection against CCHFV infection.https://journals.asm.org/doi/10.1128/mbio.03202-24Crimean-Congo hemorrhagic fever virusmonoclonal antibodybispecific antibodyimmunotherapy
spellingShingle Albert Wang
Stephanie R. Monticelli
Ariel S. Wirchnianski
Dafna M. Abelson
Ana I. Kuehne
Russell R. Bakken
Marissa Middlecamp
Michael Weingart
Olivia Vergnolle
Zachary A. Bornholdt
Crystal L. Moyer
Jacob L. Berrigan
Brandyn R. West
J. Maximilian Fels
Larry Zeitlin
Andrew S. Herbert
Kartik Chandran
Chowdhury Raihan Bikash
Jonathan R. Lai
Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus
mBio
Crimean-Congo hemorrhagic fever virus
monoclonal antibody
bispecific antibody
immunotherapy
title Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus
title_full Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus
title_fullStr Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus
title_full_unstemmed Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus
title_short Avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against Crimean-Congo hemorrhagic fever virus
title_sort avidity and variable domain spacing strongly influence the therapeutic potency of bispecific antibodies against crimean congo hemorrhagic fever virus
topic Crimean-Congo hemorrhagic fever virus
monoclonal antibody
bispecific antibody
immunotherapy
url https://journals.asm.org/doi/10.1128/mbio.03202-24
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