Site‐specific encoding of photoactivity in antibodies enables light‐mediated antibody‐antigen binding on live cells

Bridge, Thomas, Shaikh, Saher A., Thomas, Paul, Botta, Joaquin, McCormick, Peter J. and Sachdeva, Amit (2019) Site‐specific encoding of photoactivity in antibodies enables light‐mediated antibody‐antigen binding on live cells. Angewandte Chemie, 58 (50). pp. 17986-17993. ISSN 0044-8249

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Abstract

Antibodies have found applications in several fields, including, medicine, diagnostics, and nanotechnology, yet methods to modulate antibody‐antigen binding using an external agent remain limited. Here, we have developed photoactive antibody fragments by genetic site‐specific replacement of single tyrosine residues with photocaged tyrosine, in an antibody fragment, 7D12. A simple and robust assay is adopted to evaluate the light‐mediated binding of 7D12 mutants to its target, epidermal growth factor receptor (EGFR), on the surface of cancer cells. Presence of photocaged tyrosine reduces 7D12‐EGFR binding affinity by over 20‐fold in two out of three 7D12 mutants studied, and binding is restored upon exposure to 365 nm light. Molecular dynamics simulations explain the difference in effect of photocaging on 7D12‐EGFR interaction among the mutants. Finally, we demonstrate the application of photoactive antibodies in delivering fluorophores to EGFR‐positive live cancer cells in a light‐dependent manner.

Item Type:	Article
Uncontrolled Keywords:	sdg 3 - good health and well-being ,/dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being
Faculty \ School:	Faculty of Science > School of Chemistry (former - to 2024) Faculty of Science Faculty of Science > School of Pharmacy (former - to 2024)
UEA Research Groups:	Faculty of Science > Research Groups > Chemistry of Life Processes
Depositing User:	LivePure Connector
Date Deposited:	21 Oct 2019 12:30
Last Modified:	24 Sep 2024 12:51
URI:	https://ueaeprints.uea.ac.uk/id/eprint/72726
DOI:	10.1002/anie.201908655

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