For circulating tumor cells (CTCs)-based cancer diagnosis and monitoring, effective enrichment and spe-cific analysis of CTCs present significant challenges. The biomembrane interfaces can enhance the high -affinity interactions between various receptors and ligands in life activities by mediating the rearrange-ment and positioning of membrane-bound components through its fluidity. Inspired by this, we have con-structed a multivalent membrane nano-interface using aptamer-linked liposomes for the efficient capture of CTCs. Furthermore, the subsequent introduction of rolling circle amplification (RCA) reaction has in-creased the number of aptamers and extended them to the surrounding space to improve the affinity of the membrane nano-interface for CTCs. After CTCs are enriched, alkaline phosphatase overexpressed on the surface of tumor cells is used as endogenous enzyme-mediated signal amplification by catalyzing 4-nitrophenyl phosphate (pNPP) with color change, achieving the analysis of CTCs. Finally, the enrichment and visual analysis of human hepatocellular carcinoma (HepG2) with a detection limit of 10 cells/mL can be obtained by integrating the multivalent membrane nano-interface and endogenous enzyme signal amplification. The detection of the target in the serum proved this method has the potential for further clinical application and provides a potential method for studying the correlation between alkaline phos-phatase dimer and cancer progression.(c) 2022 Published by Elsevier B.V. on behalf of Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences.