Mechanistic insights into the rational design of masked antibodies
Although monoclonal antibodies have greatly improved cancer therapy, they can trigger side effects due to on-target, off-tumor toxicity. Over the past decade, strategies have emerged to successfully mask the antigen-binding site of antibodies, such that they are only activated at the relevant site,...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2022-12-01
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| Series: | mAbs |
| Subjects: | |
| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2022.2095701 |
| _version_ | 1818114709780430848 |
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| author | Carolina T. Orozco Manuela Bersellini Lorraine M. Irving Wesley W. Howard David Hargreaves Paul W. A. Devine Elise Siouve Gareth J. Browne Nicholas J. Bond Jonathan J. Phillips Peter Ravn Sophie E. Jackson |
| author_facet | Carolina T. Orozco Manuela Bersellini Lorraine M. Irving Wesley W. Howard David Hargreaves Paul W. A. Devine Elise Siouve Gareth J. Browne Nicholas J. Bond Jonathan J. Phillips Peter Ravn Sophie E. Jackson |
| author_sort | Carolina T. Orozco |
| collection | DOAJ |
| description | Although monoclonal antibodies have greatly improved cancer therapy, they can trigger side effects due to on-target, off-tumor toxicity. Over the past decade, strategies have emerged to successfully mask the antigen-binding site of antibodies, such that they are only activated at the relevant site, for example, after proteolytic cleavage. However, the methods for designing an ideal affinity-based mask and what parameters are important are not yet well understood. Here, we undertook mechanistic studies using three masks with different properties and identified four critical factors: binding site and affinity, as well as association and dissociation rate constants, which also played an important role. HDX-MS was used to identify the location of binding sites on the antibody, which were subsequently validated by obtaining a high-resolution crystal structure for one of the mask-antibody complexes. These findings will inform future designs of optimal affinity-based masks for antibodies and other therapeutic proteins. |
| first_indexed | 2024-12-11T03:55:02Z |
| format | Article |
| id | doaj.art-e42dd39c733d4c65b3014906af62224e |
| institution | Directory Open Access Journal |
| issn | 1942-0862 1942-0870 |
| language | English |
| last_indexed | 2024-12-11T03:55:02Z |
| publishDate | 2022-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj.art-e42dd39c733d4c65b3014906af62224e2022-12-22T01:21:49ZengTaylor & Francis GroupmAbs1942-08621942-08702022-12-0114110.1080/19420862.2022.2095701Mechanistic insights into the rational design of masked antibodiesCarolina T. Orozco0Manuela Bersellini1Lorraine M. Irving2Wesley W. Howard3David Hargreaves4Paul W. A. Devine5Elise Siouve6Gareth J. Browne7Nicholas J. Bond8Jonathan J. Phillips9Peter Ravn10Sophie E. Jackson11Yusuf Hamied Department of Chemistry, University of Cambridge, Cambridge, UKBiologics Engineering, R&D, AstraZeneca, Cambridge, UKBiologics Engineering, R&D, AstraZeneca, Cambridge, UKAnalytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Gaithersburg, MD, USADiscovery Sciences, R&D, AstraZeneca, Cambridge, UKAnalytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKBiologics Engineering, R&D, AstraZeneca, Cambridge, UKBiologics Engineering, R&D, AstraZeneca, Cambridge, UKAnalytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKLiving Systems Institute, University of Exeter, Exeter, UKBiologics Engineering, R&D, AstraZeneca, Cambridge, UKYusuf Hamied Department of Chemistry, University of Cambridge, Cambridge, UKAlthough monoclonal antibodies have greatly improved cancer therapy, they can trigger side effects due to on-target, off-tumor toxicity. Over the past decade, strategies have emerged to successfully mask the antigen-binding site of antibodies, such that they are only activated at the relevant site, for example, after proteolytic cleavage. However, the methods for designing an ideal affinity-based mask and what parameters are important are not yet well understood. Here, we undertook mechanistic studies using three masks with different properties and identified four critical factors: binding site and affinity, as well as association and dissociation rate constants, which also played an important role. HDX-MS was used to identify the location of binding sites on the antibody, which were subsequently validated by obtaining a high-resolution crystal structure for one of the mask-antibody complexes. These findings will inform future designs of optimal affinity-based masks for antibodies and other therapeutic proteins.https://www.tandfonline.com/doi/10.1080/19420862.2022.2095701Masked antibodiesoff-tumor cytotoxicityprotein designpro-antibodypro-drugprotein-protein interaction |
| spellingShingle | Carolina T. Orozco Manuela Bersellini Lorraine M. Irving Wesley W. Howard David Hargreaves Paul W. A. Devine Elise Siouve Gareth J. Browne Nicholas J. Bond Jonathan J. Phillips Peter Ravn Sophie E. Jackson Mechanistic insights into the rational design of masked antibodies mAbs Masked antibodies off-tumor cytotoxicity protein design pro-antibody pro-drug protein-protein interaction |
| title | Mechanistic insights into the rational design of masked antibodies |
| title_full | Mechanistic insights into the rational design of masked antibodies |
| title_fullStr | Mechanistic insights into the rational design of masked antibodies |
| title_full_unstemmed | Mechanistic insights into the rational design of masked antibodies |
| title_short | Mechanistic insights into the rational design of masked antibodies |
| title_sort | mechanistic insights into the rational design of masked antibodies |
| topic | Masked antibodies off-tumor cytotoxicity protein design pro-antibody pro-drug protein-protein interaction |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2022.2095701 |
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