Natural AIE Probe Screening and Application in Complex Samples Based on Biological Aggregation-Triggered In Situ Luminescence

Abstract The occurrence of thromboembolic diseases is closely related to the generation of biological aggregates mediated by thrombin, and the detection and regulation of its activity are crucial for research in this field. Here, an aggregate-responsive method was proposed and validated based on biological aggregation-triggered in situ luminescence, enabling the screening of natural AIE (aggregation-induced emission) probes, thrombin activity assay, and thrombin inhibitor discovery in complex samples. The results demonstrated that thrombin-induced fibrin aggregation can activate the in situ luminescence of AIE molecules in complex samples. By combining chromatography and mass spectrometry techniques, berberine was identified as a natural compound with significant AIE effects. Quantum chemical calculations simulated the molecular basis of its recognition and emission within the aggregated microenvironment. A visual sensing method for detecting thrombin activity in complex systems has been developed based on the AIE effect. This method successfully demonstrated that the extracts of taxilli herba and notoginseng radix et rhizoma contain effective ingredients that can inhibit thrombin activity. In addition, a computer-mediated virtual screening technique was developed for efficient analysis of active monomers in the candidate extracts. This study indicated that thrombin-mediated biological aggregates can be used for efficient discovery of AIE molecules in complex samples, as well as for efficient discovery of anticoagulant candidate compound.