Mammalian catechol-O-methyltransferases (COMT) are an important class of conjugative enzymes, which play a key role in the metabolism and inactivation of catechol neurotransmitters, catechol estrogens and a wide range of endobiotics and xenobiotics that bear the catechol group. Currently, COMT inhibitors are used in combination with levodopa for the treatment of Parkinson’s disease in clinical practice. The crucial role of COMT in human health has raised great interest in the development of more practical assays for highly selective and sensitive detection of COMT activity in real samples, as well as for rapid screening and characterization of COMT inhibitors as drug candidates. This review summarizes recent advances in analytical methodologies for sensing COMT activity and their applications. Several lists of biochemical assays for measuring COMT activity, including the probe substrates, along with their analytical conditions and kinetic parameters, are presented. Finally, the challenges and future perspectives in the field, such as visualization of COMT activity in vivo and in situ, are highlighted. Collectively, this review article overviews the practical assays for measuring COMT activities in complex biological samples, which will strongly facilitate the investigations on the relevance of COMT to human diseases and promote the discovery of COMT inhibitors via high-throughput screening.

哺乳动物儿茶酚-O-甲基转移酶（COMT）是一类重要的结合酶，在儿茶酚神经递质、儿茶酚雌激素以及带有儿茶酚基团的多种内生素和外源性物质的代谢和失活中发挥关键作用。目前，COMT抑制剂在临床上与左旋多巴联合用于帕金森病的治疗。 COMT 在人类健康中的关键作用引起了人们对开发更实用的检测方法的极大兴趣，这些检测方法可用于高选择性和灵敏地检测实际样品中的 COMT 活性，以及​​快速筛选和表征作为候选药物的 COMT 抑制剂。本综述总结了感测 COMT 活性的分析方法及其应用的最新进展。列出了用于测量 COMT 活性的几个生化测定方法，包括探针底物及其分析条件和动力学参数。最后，强调了该领域的挑战和未来前景，例如体内和原位 COMT 活动的可视化。总的来说，这篇综述文章概述了测量复杂生物样品中 COMT 活性的实用方法，这将极大地促进 COMT 与人类疾病相关性的研究，并促进通过高通量筛选发现 COMT 抑制剂。