For enzyme-/antibody-free and label-free biosensing, a molecularly imprinted polymer (MIP)-based membrane with phenylboronic acid (PBA) molecules, which induces the change in the density of molecular charges based on the small biomolecule–PBA diol binding, has been demonstrated to be suitable for the bioelectrical interface of biologically coupled gate field-effect transistor (bio-FET) sensors. MIP-coated gate FET sensors selectively detect various small biomolecules such as glucose, dopamine, sialic acid, and oligosaccharides without using labeled materials. In particular, the well-controlled MIP film by surface-initiated atom transfer radical polymerization (SI-ATRP) contributes to the quantitative analysis of small biomolecule sensing, resulting in potentiometric Langmuir isotherm adsorption analysis by which the parameters such as the binding affinity between small biomolecules and MIP cavities are evaluated. Also, the output electrical signal of even a random MIP-coated gate FET sensor is quantitatively analyzed using the bi-Langmuir adsorption isotherm equation, showing the adsorption mechanism of small biomolecules onto the template-specific MIP membrane. Thus, a platform based on the MIP bioelectrical interface for the bio-FET sensor is suitable for an enzyme-/antibody-free and label-free biosensing system in the fields of clinical diagnostics, drug discovery, the food industry, and environmental research.