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This work reports rapid, label-free and specific detection of the HER2 antigen using a gallium nitride (GaN) high electron mobility transistor (HEMT). Thiol-based chemistry has been utilized to immobilize the corresponding HER2 antibody in the sensing area of the sensor. The formation of a gold-sulfur complex has been confirmed through Raman spectroscopy, giving a peak at around a wavelength of 260 cm -1 . Fourier transform infrared spectroscopy and atomic force microscopy (AFM) also reveal the functionalization of thiol and free carboxylic groups. On-chip enzyme-linked immunosorbent assay has been utilized to confirm immobilization of antibody receptors on the sensing area surface, followed by current-voltage measurement. Morphology of the sensing area using AFM and electrical characterization of the sensor have been recorded before and after each functionalization process step. The sensor shows detection of the HER2 antigen in a broad range of 0.7 pg ml -1 to 10 μg ml -1 i.e. , (5 × 10 -15 to 6 × 10 -8 M). A long-time study and reusability aspect of the sensor have also been investigated that show good viability of the sensor. For the first time, a three-binding-site model based on the Langmuir isotherm has been developed for HER2 detection using GaN-HEMTs with three dissociation constants, i.e. , 7 × 10 -10 , 8.8 × 10 -11 , and 7.2 × 10 -9 M, respectively.