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Biomarker expression both on the cell surface and in serum is directly related to the pathological process of tumor. Based on the interaction between the ligand and the protein receptor, a label-free photoelectrochemical (PEC) biosensing interface with good antifouling ability was proposed for tumor cell detection. TiO 2 nanotube (NT) arrays were used as the substrate to enhance the ability of the biosensor to capture the target. Mercapto-terminated 8-arm poly(ethylene glycol) was introduced onto the electrode surface by the deposition of Au nanoparticles on TiO 2 NTs, creating an antifouling molecular layer. The recognition ligand hyaluronic acid (HA) was functionalized by dopamine and introduced onto the sensing surface based on the unique chelating interaction between the catechol group and the titanium atom. Benefitting from the specific recognition of HA with CD44 and the 3D porous structures of NTs, the constructed PEC biosensor showed excellent abilities toward the detection of MDA-MB-231 breast tumor cells and the soluble form of CD44. The ligand-receptor PEC sensing strategy has promising potential for the detection of tumor cells and protein biomarkers.