Cargando…
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...
Autores principales: | , , , , , , , , , , , , |
---|---|
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 |
_version_ | 1785059165688823808 |
---|---|
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. |
format | Online Article Text |
id | pubmed-10269415 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT johanssonmariau designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT weinertchristopher designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT reichardtdietrichalexander designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT mahlerdana designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT diemdania designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT hesschristian designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT feusidiana designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT carnalsimon designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT tietzjulia designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT giezendannernoreen designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT spigafabiomario designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT urechdavid designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies AT warmuthstefan designofantibodyvariablefragmentswithreducedreactivitytopreexistingantidrugantibodies |