Photoactivatable Fluorescent Tags for Dual-Modality Positron Emission Tomography Optical Imaging

Fluorescent protein conjugates are vital tools in a wide range of scientific disciplines from basic biochemical research to applications in clinical pathology and intraoperative surgery. We report the synthesis and characterization of photoactivatable fluorophores (PhotoTags) based on the functionalization of coumarin, fluorescein, BODIPY, rhodamine B, and cyanine dyes with a photochemically active aryl azide group. Photochemical labeling experiments using human serum albumin produced fluorescent proteins in high yields under irradiation with ultraviolet light for <15 min. We also synthesized DFO-RhodB-PEG₃-ArN₃─a photoactivatable compound that can be radiolabeled with ⁸⁹Zr for applications in optical imaging and positron emission tomography. One-pot ⁸⁹Zr-radiolabeling and light-induced protein conjugation produced [⁸⁹Zr]ZrDFO-RhodB-PEG₃-azepin-trastuzumab. Proof-of-concept studies in vitro and in vivo confirmed that [⁸⁹Zr]ZrDFO-RhodB-PEG₃-azepin-trastuzumab is a potential dual-modality agent for detecting human epidermal growth factor receptor 2 (HER2/neu) expression. Overall, the PhotoTag technology represents a rapid, synthetically versatile, and user-friendly approach for generating novel protein conjugates.