Activity-Triggered Chemiluminescent Probe for α-L-Fucosidase Detection from In Vitro to In Vivo

Hepatocellular carcinoma (HCC), one of the most prevalent malignancies with high mortality, necessitates early intervention and prognostic monitoring to improve patient survival. However, the lack of ideal detection methods remains a critical challenge. α-L-Fucosidase (AFU), a key lysosomal hydrolase, has emerged as a promising biomarker for early-stage hepatocellular carcinoma diagnosis. Herein, we developed a chemiluminescent probe (AFU-CL) capable of specifically responding to AFU. AFU-CL enables rapid and highly selective AFU detection, exhibiting exceptional sensitivity (limit of detection = 4.87 × 10–4 mU/mL) and superior signal-to-noise ratios compared to conventional fluorescent probes. At the cellular level, AFU-CL enabled real-time visualization of endogenous AFU activity in cultured HCC cells (MHCC-97H), demonstrating distinct signal patterns between malignant and normal hepatocytes (MIHA). Progressing to in vivo imaging, the probe successfully achieved in situ visualization of tumor sites in HCC-bearing mouse models, with its specific chemiluminescent signals clearly distinguishing tumor tissues from normal tissues. Most significantly, this detection capability was effectively translated to clinical applications, as AFU-CL-based analysis reliably differentiated serum samples from HCC patients and healthy controls, confirming the probe’s potential for clinical diagnosis. This strategy provides a promising platform for early HCC diagnosis and elucidating the functional role of AFU in cancer progression.