Tools
ToolsBridge, Thomas, Wegmann, Udo, Crack, Jason C., Orman, Kate, Shaikh, Saher A., Farndon, William, Martins, Carlo, Saalbach, Gerhard and Sachdeva, Amit (2023) Site-specific encoding of photoactivity and photoreactivity into antibody fragments. Nature Chemical Biology, 19 (6). 740–749. ISSN 1552-4450
Preview |
PDF (s41589-022-01251-9)
- Published Version
Available under License Creative Commons Attribution. Download (15MB) | Preview |
Abstract
Design of biomolecules that perform two or more distinct functions in response to light remains challenging. Here, we have introduced concurrent photoactivity and photoreactivity into an epidermal growth factor receptor (EGFR)-targeting antibody fragment, 7D12. This was achieved by site-specific incorporation of photocaged tyrosine (pcY) for photoactivity and p-benzoyl-ʟ-phenylalanine (Bpa) for photoreactivity into 7D12. We identified a position for installing Bpa in 7D12 that has minimal effect on 7D12–EGFR binding affinity in the absence of light. Upon exposure to 365-nm light, this Bpa-containing 7D12 mutant forms a covalent bond with EGFR in an antigen-specific manner. We then developed a method for site-specific incorporation of pcY and Bpa at two distinct sites in 7D12. Finally, we demonstrated that in the absence of light, this pcY- and Bpa-containing mutant of 7D12 does not bind to EGFR, but irradiation with 365-nm light activates (1) specific binding and (2) covalent bond formation with EGFR.
| Item Type: | Article |
|---|---|
| Additional Information: | Author acknowledgements: We thank P. G. Schultz, S. Balasubramanian and J. McCafferty for sharing plasmids with Addgene and thank Addgene for providing the plasmids. We also thank J. Chin for sharing plasmids from his laboratory and N. Le Brun for providing access to the mass spectrometer. Funding Information: This work was funded by the Wellcome Trust (204593/Z/16/Z), BBSRC (BB/R004692/1) and the University of East Anglia. |
| Uncontrolled Keywords: | molecular biology,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1312 |
| Faculty \ School: | Faculty of Science > School of Chemistry (former - to 2024) |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 21 Aug 2023 08:30 |
| Last Modified: | 16 Dec 2024 01:40 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/92892 |
| DOI: | 10.1038/s41589-022-01251-9 |
Downloads
Downloads per month over past year
Actions (login required)
 |
View Item |