Host-guest recognition plays an important role in biological analysis and biosensing. Accordingly, great efforts have been devoted to the development of sensors using versatile 3D surface materials. These functionalized nanomaterials possess the advantages of high selectivity and visual signals, enabling the selective detection of ions, amino acids, proteins, and other biological molecules. Therefore, in this feature article, we present some significant and representative examples of molecular recognition on calixarene functionalized 3D surface nanomaterials. By virtue of host-guest interactions, the functional interfaces displaying high selectivity and featuring a reversible switch response towards the environmental stimuli with various signal output (electrochemical and optical signals) are herein introduced. Furthermore, various 3D surface materials with unique signal amplification in molecular recognition are presented, including quantum dots (QDs), metal nanoparticles (NPs), nanotubes, and mesoporous silica. These excellent properties enable calixarene-functionalized 3D materials to become an outstanding platform for molecular recognition, offering convenient approaches for sensing and separation.