{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T09:48:03Z","timestamp":1765964883303,"version":"3.48.0"},"reference-count":46,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T00:00:00Z","timestamp":1765929600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001659","name":"German Research Foundation","doi-asserted-by":"publisher","award":["WO 2178\/3\u20131"],"award-info":[{"award-number":["WO 2178\/3\u20131"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Federal Ministry of Economic Affairs and Climate Action","award":["03THW15H04"],"award-info":[{"award-number":["03THW15H04"]}]},{"name":"Federal Ministry of Research, Technology and Space","award":["03VP12060"],"award-info":[{"award-number":["03VP12060"]}]},{"name":"Open Access Publication Fund of the University of Freiburg"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Background: Non-muscle invasive bladder cancer (NMIBC) has limited therapeutic options and high recurrence rates. Photoimmunotherapy (PIT) enables targeted tumor ablation using antibody-photosensitizer conjugates and light activation. We evaluated EGFR, Nectin-4, and TROP-2 as PIT targets using cysteine-modified antibodies conjugated to the photosensitizer WB692-CB2. Methods: Antibodies derived from Cetuximab (Cmb, anti-EGFR), Enfortumab (Enf, anti-Nectin-4), and Sacituzumab (Sac, anti-TROP-2) were engineered with T120C and D265C mutations in the heavy chains for site-specific dye conjugation. Binding of the conjugates to BC cells was tested by flow cytometry and light-induced cytotoxicity of the conjugates, alone or in combination, was assessed by viability assays following irradiation. Results: Cysteine-modified antibodies were produced as intact IgG molecules and were efficiently conjugated with WB692-CB2 without loss of antigen specificity. SacT120C\/D265C-WB692-CB2 showed the highest target binding and achieved near-complete cell killing at a red-light dose of 32 J\/cm2. CmbT120C\/D265C-WB692-CB2 required a fourfold higher light dose for comparable efficacy, while EnfT120C\/D265C-WB692-CB2 demonstrated lower potency. No cytotoxicity was observed in antigen-negative cells. Combined treatment enhanced cytotoxicity, indicating additive phototherapeutic effects. Conclusions: Our findings suggest that PIT targeting EGFR, Nectin-4, or TROP-2 merits further preclinical development as a targeted therapeutic approach for NMIBC, including potential combinatorial or personalized strategies.<\/jats:p>","DOI":"10.3390\/molecules30244802","type":"journal-article","created":{"date-parts":[[2025,12,17]],"date-time":"2025-12-17T09:43:46Z","timestamp":1765964626000},"page":"4802","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Exploring EGFR, Nectin-4, and TROP-2 as Therapeutic Targets for Bladder Cancer Photoimmunotherapy"],"prefix":"10.3390","volume":"30","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6462-9778","authenticated-orcid":false,"given":"Isis","family":"Wolf","sequence":"first","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nora","family":"Giess","sequence":"additional","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C\u00e9line","family":"Roider","sequence":"additional","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susanne","family":"Schultze-Seemann","sequence":"additional","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonas","family":"Storz","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry, Albert-Ludwigs-Universit\u00e4t Freiburg, Albertstr. 21, 79104 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel B.","family":"Werz","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry, Albert-Ludwigs-Universit\u00e4t Freiburg, Albertstr. 21, 79104 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arkadiusz","family":"Miernik","sequence":"additional","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Gratzke","sequence":"additional","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6779-4325","authenticated-orcid":false,"given":"Philipp","family":"Wolf","sequence":"additional","affiliation":[{"name":"Department of Urology, Medical Center-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany"},{"name":"Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,17]]},"reference":[{"key":"ref_1","first-page":"209","article-title":"Global cancer statistics 2020: Globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. 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