Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies
ABSTRACTUpon 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 previou...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2023-12-01
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| Series: | mAbs |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2023.2215887 |
| _version_ | 1797356288124911616 |
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| author | Maria U. Johansson Christopher Weinert Dietrich Alexander Reichardt Dana Mahler Dania Diem Christian Hess Diana Feusi Simon Carnal Julia Tietz Noreen Giezendanner Fabio Mario Spiga David Urech Stefan Warmuth |
| author_facet | Maria U. Johansson Christopher Weinert Dietrich Alexander Reichardt Dana Mahler Dania Diem Christian Hess Diana Feusi Simon Carnal Julia Tietz Noreen Giezendanner Fabio Mario Spiga David Urech Stefan Warmuth |
| author_sort | Maria U. Johansson |
| collection | DOAJ |
| description | ABSTRACTUpon 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. |
| first_indexed | 2024-03-08T14:24:27Z |
| format | Article |
| id | doaj.art-b59c6c9046354f519ced556acc07494f |
| institution | Directory Open Access Journal |
| issn | 1942-0862 1942-0870 |
| language | English |
| last_indexed | 2024-03-08T14:24:27Z |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj.art-b59c6c9046354f519ced556acc07494f2024-01-13T11:27:51ZengTaylor & Francis GroupmAbs1942-08621942-08702023-12-0115110.1080/19420862.2023.2215887Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodiesMaria U. Johansson0Christopher Weinert1Dietrich Alexander Reichardt2Dana Mahler3Dania Diem4Christian Hess5Diana Feusi6Simon Carnal7Julia Tietz8Noreen Giezendanner9Fabio Mario Spiga10David Urech11Stefan Warmuth12Numab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandNumab Therapeutics AG, Horgen, SwitzerlandABSTRACTUpon 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.https://www.tandfonline.com/doi/10.1080/19420862.2023.2215887Anti-drug antibodiesantibodyantibody fragmentbiotherapeuticsimmunogenicitymultispecific |
| spellingShingle | Maria U. Johansson Christopher Weinert Dietrich Alexander Reichardt Dana Mahler Dania Diem Christian Hess Diana Feusi Simon Carnal Julia Tietz Noreen Giezendanner Fabio Mario Spiga David Urech Stefan Warmuth Design of antibody variable fragments with reduced reactivity to preexisting anti-drug antibodies mAbs Anti-drug antibodies antibody antibody fragment biotherapeutics immunogenicity multispecific |
| 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 | Anti-drug antibodies antibody antibody fragment biotherapeutics immunogenicity multispecific |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2023.2215887 |
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