High-throughput optical assays for sensing serine hydrolases in living systems and their applications

Serine hydrolases (SHs) comprise a large family of hydrolytic enzymes that play crucial roles in numerous physiological and pathological processes, such as blood coagulation, cancer, inflammation, and diabetes. This review summarizes the newest advances in the design and development of specific optical substrates for mammalian SHs and related high-throughput optical assays for sensing target SHs in living systems, as well as their applications in bioanalytical and biomedical fields. The design principles and the response modes for SHs optical substrates, as well as the structural information and kinetic parameters of reported probes for mammalian SHs, have been well summarized. Furthermore, the biomedical applications of SHs optical substrates in various fields, including sensing and imaging analysis of target enzyme in biological systems, high-throughput screening of enzyme inhibitors, have been demonstrated. Finally, the challenges and future perspectives in this field are also highlighted.