手性药物的对映异构体在人体内具有不同的药理毒理效应，手性药物的分离具有重要的现实意义。作为内源性手性选择剂，酶分子种类繁多，依托酶类手性固定相的毛细管电色谱技术可实现更为广谱手性药物的快速高效分离。然而，毛细管柱内酶固定化效率不高且易失活，成为酶类手性选择剂应用的难点问题。本项目拟在前期工作基础上，借助自主研发的氧化石墨烯（GO）修饰毛细管整体柱，构建性能稳定的酶固载化微环境，充分发挥GO对酶手性分离能力的提升作用，获得高柱效高分离度的手性分离电色谱图。同时，利用胰蛋白酶（TRYP）与糜蛋白酶（CHT）手性识别效能的互补性，建立基于双酶联用策略的多种类手性药物分离新方法。此外，将该体系与前沿分析法结合，从分子层面探讨各单旋体与TRYP/CHT相互作用对手性分离的影响。此项目有望为实现酶类手性选择剂高效快速广谱的分离性能提供新的技术手段，并为体内酶-手性药物相互作用机理研究提供参考。

The living systems present distinctive physiological responses to different enantiomeric forms of a single drug, chiral separation exhibits its significant importance nowadays. Enzymes is a type of endogenous chiral selector, and enzyme-based capillary electrochromatography technique can be used to separate different enantiomers with high performance due to the wide varieties of enzymes. However, the application of enzyme-involved chiral selectors is limited since the activity and immobilization capacity of enzymes are easily affected by the immobilization process in capillaries. Following our previous works, a graphene oxide-modified capillary monolith can be applied to build a stable microenvironment for enzyme immobilization and achieve chiral separation with high column efficiency and resolution. Furthermore, for the sake of separating different types of enantiomers, bi-enzymatic system will be developed to combine the enantiomeric separation ability of Trypsin (TRYP) and Chymotrypsin (CHT). In addition, in order to investigate the chiral separation process at a molecular level, frontal analysis technology will also be applied to discuss the interaction mechanism between TRYP/CHT and enantiomers. This project is expected to provide a novel technique for the development of enzyme-based chiral separation systems with high efficiency and broad applicability. Meanwhile, this technology can be utilized to discuss enzyme-enantiomers interaction mechanism.