As a common substance in the human body, melibiose has a good effect on treating inflammation, and chiral selectively assembled behaviors often accompany this metabolic activity. Therefore, studies on melibiose's chiral selectively assembled behaviors arouse broad interest, making it better for us to understand similar bio-processes. Herein, we modified (R)/(S)-phenylethylamine calix[4]arenes onto the gold surface to analyze the chiral selective behaviors for melibiose. Contact angle and electrochemical experimental results showed that this functional surface modified with (S)-phenylethylamine calix[4]arene achieved selective adsorption of melibiose in the condition of pH = 7 based on the weak host–guest interaction. Furthermore, these functional surfaces have been realized by amplifying the weak selective interaction, and these chiral selective sensors have been constructed for the melibiose.