In nature, cellular molecule sensing is usually achieved at the environment/membrane interface. In the meantime, rapid growth of nanotechnology is increasingly pushing engineered nanomaterials to interact with biological surfaces. Herein, inspired by trans-membrane signal transduction, a nano-bio interface has been constructed in this work for biosensing application. The interface is formed between a selective biomembrane mimetic surface (SBMMS) and a function-oriented 2D nanohybrid. Based on the design, target recognition can be performed in a biologically favorable environment, and the nano-bio interaction can be transduced into amplified electrochemical readouts. Furthermore, this sensing platform can be used to analyze various kinds of targets, including proteins, nucleic acids, and small molecules, just by changing the biorecognition element. Low detection limits and wide detection ranges can also be obtained. So, this nano-bio interface may provide a new platform for bioanalytical research in the future.