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<p>Developing fluorescent probes with high selectivity for target proteins in living cells remains a significant challenge. Conventional probes, such as nitrobenzoxadiazole (NBD) derivatives, often suffer from limited selectivity and short emission wavelengths, limiting their performance in complex biological environments. Here, we report the design and synthesis of NBSe-biotin, a red-emitting fluorescent probe based on the nitrobenzoselenadiazole (NBSe) scaffold. Compared to NBD analogues, NBSe-biotin exhibits red-shifted absorption and emission, enhanced chemical selectivity, and a turn-on fluorescence response upon specific binding to avidin, with emission at 604 nm. Live-cell imaging demonstrates that NBSe-biotin selectively visualizes overexpressed biotin receptors on HeLa cell membranes, with minimal background fluorescence in 293T control cells. Beyond imaging, this strategy enables the covalent attachment of small-molecule photosensitizers to proteins, and the receptor-mediated activation of NBSe-biotin phototoxicity in live cells illustrates its potential to incorporate diverse cancer biomarker-targeting ligands. Together, these features establish NBSe-biotin as a versatile platform for fluorescence-guided protein detection, live-cell imaging, and targeted phototherapy.</p>

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An Activatable Nitrobenzoselenadiazole-based Probe for Selective Protein Labeling and Targeted Phototherapy

  • 期刊论文

  • 发表日期：

    2026年3月17日

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  • Xiaojie Ren,
  • Wenlong Yi,
  • Li Yang,
  • Xinyu Ye,
  • Cheuk Hin Li,
  • Fan Zhang,
  • Jiafu Lin,
  • Hongyan Sun

摘要

Developing fluorescent probes with high selectivity for target proteins in living cells remains a significant challenge. Conventional probes, such as nitrobenzoxadiazole (NBD) derivatives, often suffer from limited selectivity and short emission wavelengths, limiting their performance in complex biological environments. Here, we report the design and synthesis of NBSe-biotin, a red-emitting fluorescent probe based on the nitrobenzoselenadiazole (NBSe) scaffold. Compared to NBD analogues, NBSe-biotin exhibits red-shifted absorption and emission, enhanced chemical selectivity, and a turn-on fluorescence response upon specific binding to avidin, with emission at 604 nm. Live-cell imaging demonstrates that NBSe-biotin selectively visualizes overexpressed biotin receptors on HeLa cell membranes, with minimal background fluorescence in 293T control cells. Beyond imaging, this strategy enables the covalent attachment of small-molecule photosensitizers to proteins, and the receptor-mediated activation of NBSe-biotin phototoxicity in live cells illustrates its potential to incorporate diverse cancer biomarker-targeting ligands. Together, these features establish NBSe-biotin as a versatile platform for fluorescence-guided protein detection, live-cell imaging, and targeted phototherapy.