A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs
Engineered antibody formats, such as antibody fragments and bispecifics, have the potential to offer improved therapeutic efficacy compared to traditional full-length monoclonal antibodies (mAbs). However, the translation of these non-natural molecules into successful therapeutics can be hampered by...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2024.2403156 |
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| author | Itzel Condado-Morales Fabian Dingfelder Isabel Waibel Oliver M. Turnbull Bhargav Patel Zheng Cao Jais Rose Bjelke Susanne Nedergaard Grell Anja Bennet Alissa M. Hummer Matthew I. J. Raybould Charlotte M. Deane Thomas Egebjerg Nikolai Lorenzen Paolo Arosio |
| author_facet | Itzel Condado-Morales Fabian Dingfelder Isabel Waibel Oliver M. Turnbull Bhargav Patel Zheng Cao Jais Rose Bjelke Susanne Nedergaard Grell Anja Bennet Alissa M. Hummer Matthew I. J. Raybould Charlotte M. Deane Thomas Egebjerg Nikolai Lorenzen Paolo Arosio |
| author_sort | Itzel Condado-Morales |
| collection | DOAJ |
| description | Engineered antibody formats, such as antibody fragments and bispecifics, have the potential to offer improved therapeutic efficacy compared to traditional full-length monoclonal antibodies (mAbs). However, the translation of these non-natural molecules into successful therapeutics can be hampered by developability challenges. Here, we systematically analyzed 64 different antibody constructs targeting Tumor Necrosis Factor (TNF) which cover 8 distinct molecular format families, encompassing full-length antibodies, various types of single chain variable fragments, and bispecifics. We measured 15 biophysical properties related to activity, manufacturing, and stability, scoring variants with a flag-based risk approach and a recent in silico developability profiler. Our comparative assessment revealed that overall developability is higher for the natural full-length antibody format. Bispecific antibodies, antibodies with scFv fragments at the C-terminus of the light chain, and single-chain Fv antibody fragments (scFvs) have intermediate developability properties, while more complicated formats, such as scFv- scFv, bispecific mAbs with one Fab exchanged with a scFv, and diabody formats are collectively more challenging. In particular, our study highlights the propensity for fragmentation and aggregation, both in bulk and at interfaces, for many current engineered formats. |
| format | Article |
| id | doaj-art-902ae4a0e4394deeb9b5cd245a0a3c88 |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-902ae4a0e4394deeb9b5cd245a0a3c882025-01-31T04:19:38ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2403156A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructsItzel Condado-Morales0Fabian Dingfelder1Isabel Waibel2Oliver M. Turnbull3Bhargav Patel4Zheng Cao5Jais Rose Bjelke6Susanne Nedergaard Grell7Anja Bennet8Alissa M. Hummer9Matthew I. J. Raybould10Charlotte M. Deane11Thomas Egebjerg12Nikolai Lorenzen13Paolo Arosio14Department of Biophysics and Injectable Formulation, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Biophysics and Injectable Formulation, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, Swiss Federal Institute of Technology, Zurich, SwitzerlandDepartment of Statistics, University of Oxford, Oxford, UKDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, Swiss Federal Institute of Technology, Zurich, SwitzerlandDepartment of Bioanalysis, Beijing Novo Nordisk Pharmaceutical Science & Technology Co. Ltd (Novo Nordisk R&D China), Beijing, ChinaDepartment of Purification Technologies, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Kidney Biology, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Kidney Biology, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Statistics, University of Oxford, Oxford, UKDepartment of Statistics, University of Oxford, Oxford, UKDepartment of Statistics, University of Oxford, Oxford, UKDepartment of Mammalian Expression, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Biophysics and Injectable Formulation, Global Research Technologies, Novo Nordisk A/S, Måløv, DenmarkDepartment of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, Swiss Federal Institute of Technology, Zurich, SwitzerlandEngineered antibody formats, such as antibody fragments and bispecifics, have the potential to offer improved therapeutic efficacy compared to traditional full-length monoclonal antibodies (mAbs). However, the translation of these non-natural molecules into successful therapeutics can be hampered by developability challenges. Here, we systematically analyzed 64 different antibody constructs targeting Tumor Necrosis Factor (TNF) which cover 8 distinct molecular format families, encompassing full-length antibodies, various types of single chain variable fragments, and bispecifics. We measured 15 biophysical properties related to activity, manufacturing, and stability, scoring variants with a flag-based risk approach and a recent in silico developability profiler. Our comparative assessment revealed that overall developability is higher for the natural full-length antibody format. Bispecific antibodies, antibodies with scFv fragments at the C-terminus of the light chain, and single-chain Fv antibody fragments (scFvs) have intermediate developability properties, while more complicated formats, such as scFv- scFv, bispecific mAbs with one Fab exchanged with a scFv, and diabody formats are collectively more challenging. In particular, our study highlights the propensity for fragmentation and aggregation, both in bulk and at interfaces, for many current engineered formats.https://www.tandfonline.com/doi/10.1080/19420862.2024.2403156AntibodybispecificsfragmentsdevelopabilityaggregationFragmentation |
| spellingShingle | Itzel Condado-Morales Fabian Dingfelder Isabel Waibel Oliver M. Turnbull Bhargav Patel Zheng Cao Jais Rose Bjelke Susanne Nedergaard Grell Anja Bennet Alissa M. Hummer Matthew I. J. Raybould Charlotte M. Deane Thomas Egebjerg Nikolai Lorenzen Paolo Arosio A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs mAbs Antibody bispecifics fragments developability aggregation Fragmentation |
| title | A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs |
| title_full | A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs |
| title_fullStr | A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs |
| title_full_unstemmed | A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs |
| title_short | A comparative study of the developability of full-length antibodies, fragments, and bispecific formats reveals higher stability risks for engineered constructs |
| title_sort | comparative study of the developability of full length antibodies fragments and bispecific formats reveals higher stability risks for engineered constructs |
| topic | Antibody bispecifics fragments developability aggregation Fragmentation |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2403156 |
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