ABSTRACT Protein aggregation is an important pathological feature of cardiovascular disease. Therein, thrombin‐mediated fibrin aggregation is one of the mechanisms of thrombosis, accurate monitoring of which is significant for the research of thrombosis. In this study, the optical properties and theoretical calculations confirmed that the AIE probe could specifically illuminate fibrin aggregates without interference, and its signal response was positively correlated with thrombin activity. Therefore, the biosensing technique can realize in situ monitoring of the coagulation process and rapid identification of active substances in complex systems. Furthermore, to detect anticoagulant active monomers, a biosensing targeted affinity screening (BioSTAS) technology was established by combining the above biosensing technique with the affinity chromatography technique. The rapid identification of active substances was achieved through biosensing, and then active monomers were captured by affinity chromatography. As a result, two agents with anticoagulant activity, rhein and oleanolic acid, were discovered via the screening of more than 30 kinds of natural products and commercial preparations. This study not only provides a new idea for the application of AIE probes in the dynamic monitoring of protein aggregation but also establishes an innovative strategy for screening active agents from a complex system through the integration of biosensing and affinity chromatography technologies.
Zhang et al. (2026) studied this question.