To construct an electrochemical biosensing platform, we propose a glucose sensor whose electrode interface was modified by mesoporous silica (MPSi) as an electronic signal transmission interface between a biomarker and an electrochemical device. We develop an enzyme-free glucose sensor using an MPSicoated Ta 2 O 5 electrode in an actual biological fluid such as blood serum. MPSi includes a phenylboronic acid (PBA) molecule, in which glucose binds to a synthesized PBA−silane compound in an ca. 150 nm thick MPSi nanolayer, which changes the density of molecular charges of the PBA/glucose complex on the surface of MPSi. The charge changes derived from the equilibrium reaction of PBA with glucose lead to changes in surface potential of the Ta 2 O 5 electrode, and the surface potential changes depending on glucose concentration were measured by a potentiometric detector. As a result, a remarkable surface potential response was observed in the vicinity of neutral pH. K d = 6.0 mM and V max = 194 mV were obtained from the fitting curve of the Langmuir adsorption isotherm. Finally, we confirmed the glucose response of the PBA−MPSi-coated Ta 2 O 5 substrate in human serum by considering the influence of various contaminants. Although the surface potential change was suppressed by approximately one-third of that in the buffer system, it was suggested that it could be applied to measurements in the blood glucose concentration range. From the results of this study, it was clarified that blood-level glucose response could be monitored using a PBA−MPSi-coated Ta 2 O 5 substrate, which suggests the possibility of using a nonenzymatic glucose sensor as an alternative to the existing enzyme sensor.