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Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies

Upon reformatting of an antibody to single-chain variable fragment format, a region in the former variable/constant domain interface of the heavy chain becomes accessible for preexisting (PE) anti-drug antibody (ADA) binding. The region exposed because of this reformatting contains a previously hidd...

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Autores principales: Johansson, Maria U., Weinert, Christopher, Reichardt, Dietrich Alexander, Mahler, Dana, Diem, Dania, Hess, Christian, Feusi, Diana, Carnal, Simon, Tietz, Julia, Giezendanner, Noreen, Spiga, Fabio Mario, Urech, David, Warmuth, Stefan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10269415/
https://www.ncbi.nlm.nih.gov/pubmed/37312434
http://dx.doi.org/10.1080/19420862.2023.2215887
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author Johansson, Maria U.
Weinert, Christopher
Reichardt, Dietrich Alexander
Mahler, Dana
Diem, Dania
Hess, Christian
Feusi, Diana
Carnal, Simon
Tietz, Julia
Giezendanner, Noreen
Spiga, Fabio Mario
Urech, David
Warmuth, Stefan
author_facet Johansson, Maria U.
Weinert, Christopher
Reichardt, Dietrich Alexander
Mahler, Dana
Diem, Dania
Hess, Christian
Feusi, Diana
Carnal, Simon
Tietz, Julia
Giezendanner, Noreen
Spiga, Fabio Mario
Urech, David
Warmuth, Stefan
author_sort Johansson, Maria U.
collection PubMed
description Upon reformatting of an antibody to single-chain variable fragment format, a region in the former variable/constant domain interface of the heavy chain becomes accessible for preexisting (PE) anti-drug antibody (ADA) binding. The region exposed because of this reformatting contains a previously hidden hydrophobic patch. In this study, mutations are introduced in this region to reduce PE ADA reactivity and concomitantly reduce the hydrophobic patch. To enhance our understanding of the importance of individual residues in this region with respect to PE ADA reactivity, a total of 50 molecules for each of two antibodies against different tumor-associated antigens were designed, produced, and characterized by an arsenal of biophysical methods. The aim was to identify suitable mutations that reduce, or completely eliminate, PE ADA reactivity to variable fragments, without compromising biophysical and pharmacodynamic properties. Computational methods were used to pinpoint key residues to mutate and to evaluate designed molecules in silico, in order to reduce the number of molecules to produce and characterize experimentally. Mutation of two threonine residues, Thr101 and Thr146 in the variable heavy domain, proved to be critical to eliminate PE ADA reactivity. This may have important implications in optimizing early drug development for antibody fragment-based therapeutics.
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spelling pubmed-102694152023-06-16 Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies Johansson, Maria U. Weinert, Christopher Reichardt, Dietrich Alexander Mahler, Dana Diem, Dania Hess, Christian Feusi, Diana Carnal, Simon Tietz, Julia Giezendanner, Noreen Spiga, Fabio Mario Urech, David Warmuth, Stefan MAbs Report Upon reformatting of an antibody to single-chain variable fragment format, a region in the former variable/constant domain interface of the heavy chain becomes accessible for preexisting (PE) anti-drug antibody (ADA) binding. The region exposed because of this reformatting contains a previously hidden hydrophobic patch. In this study, mutations are introduced in this region to reduce PE ADA reactivity and concomitantly reduce the hydrophobic patch. To enhance our understanding of the importance of individual residues in this region with respect to PE ADA reactivity, a total of 50 molecules for each of two antibodies against different tumor-associated antigens were designed, produced, and characterized by an arsenal of biophysical methods. The aim was to identify suitable mutations that reduce, or completely eliminate, PE ADA reactivity to variable fragments, without compromising biophysical and pharmacodynamic properties. Computational methods were used to pinpoint key residues to mutate and to evaluate designed molecules in silico, in order to reduce the number of molecules to produce and characterize experimentally. Mutation of two threonine residues, Thr101 and Thr146 in the variable heavy domain, proved to be critical to eliminate PE ADA reactivity. This may have important implications in optimizing early drug development for antibody fragment-based therapeutics. Taylor & Francis 2023-06-13 /pmc/articles/PMC10269415/ /pubmed/37312434 http://dx.doi.org/10.1080/19420862.2023.2215887 Text en © 2023 Numab Therapeutics AG. Published with license by Taylor & Francis Group, LLC. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) ), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.
spellingShingle Report
Johansson, Maria U.
Weinert, Christopher
Reichardt, Dietrich Alexander
Mahler, Dana
Diem, Dania
Hess, Christian
Feusi, Diana
Carnal, Simon
Tietz, Julia
Giezendanner, Noreen
Spiga, Fabio Mario
Urech, David
Warmuth, Stefan
Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
title Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
title_full Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
title_fullStr Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
title_full_unstemmed Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
title_short Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
title_sort design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10269415/
https://www.ncbi.nlm.nih.gov/pubmed/37312434
http://dx.doi.org/10.1080/19420862.2023.2215887
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