| Autor: |
Orozco CT; Yusuf Hamied Department of Chemistry, University of Cambridge, Cambridge, UK.; Biologics Engineering, R&D, AstraZeneca, Cambridge, UK.; Analytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UK., Bersellini M; Biologics Engineering, R&D, AstraZeneca, Cambridge, UK., Irving LM; Biologics Engineering, R&D, AstraZeneca, Cambridge, UK., Howard WW; Analytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Gaithersburg, MD, USA., Hargreaves D; Discovery Sciences, R&D, AstraZeneca, Cambridge, UK., Devine PWA; Analytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UK., Siouve E; Biologics Engineering, R&D, AstraZeneca, Cambridge, UK.; Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, UK., Browne GJ; Biologics Engineering, R&D, AstraZeneca, Cambridge, UK., Bond NJ; Analytical Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UK., Phillips JJ; Living Systems Institute, University of Exeter, Exeter, UK., Ravn P; Biologics Engineering, R&D, AstraZeneca, Cambridge, UK.; Department of Biotherapeutic Discovery, H. Lundbeck A/S, Copenhagen, Denmark., Jackson SE; Yusuf Hamied Department of Chemistry, University of Cambridge, Cambridge, UK. |
| Abstrakt: |
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. |
