To rapidly and accurately screen compounds present in traditional Chinese herbal medicines for α-glycosidase inhibitors, we synthesized a novel nano-affinity material to enable the targeted screening and enrichment of α-glycosidase inhibitors. This nano-affinity material was synthesized by self-assembly involving α-glycosidase and Cu2+. A systematic analysis revealed the flower-like structure of this nano-material, with a diameter of 7.564 μm. The highest enzyme activity was 214.03% compared to free enzymes. This novel material was used to screen and enrich α-glycosidase inhibitors from Chinese rhubarb. Ten compounds were detected. Of these, l-epicatechin and anthraquinone displayed inhibitory activity. This novel nano-hybrid affinity material allows the rapid screening and separation of active components in complex systems. [ABSTRACT FROM AUTHOR]