An integrated dual-phase biosensing and ligand fishing platform for high-throughput screening of HAase inhibitors and clinical urine analysis

A new bladder cancer diagnosis and HAase inhibitor screening platform was developed. • Biosensor can detect HAase in urine and distinguish patients without preprocessing. • This method shows great potential for rapid screening of HAase inhibitor drugs. • Cinnamon Bark exhibited a significant HAase inhibition effect among 56 herbs. • Cinnamaldehyde was identified as a HAase inhibitor with an IC₅₀ of 396.5 μg/mL. Hyaluronidase (HAase) is a potent urinary biomarker for bladder cancer, yet its clinical detection and the discovery of related inhibitors are often hindered by complex procedures and susceptibility to interference. To address these challenges, this study developed an integrated strategy that combines a dual-phase biosensor with immobilized HAase-based ligand fishing. The HAase-responsive biosensor enables both high-throughput screening of inhibitors from herbal extracts and direct, pretreatment-free analysis of clinical urine samples, effectively distinguishing bladder cancer patients from healthy individuals. Meanwhile, the complementary ligand fishing module allows precise capture and identification of bioactive compounds from complex mixtures, leading to the discovery of cinnamaldehyde as a candidate inhibitor (IC 50  = 396.5 μg/mL). In conclusion, this platform provides a versatile strategy that not only facilitates the rapid discovery of HAase-targeting agents from natural products but also offers a simple and interference-resistant approach for non-invasive monitoring of bladder cancer. Download: Download high-res image (384KB) Download: Download full-size image