Screen-printed carbon electrode (SPCE) is a useful technique that has been widely used in the practical application of biosensors oriented to point-of-care testing (POCT) due to its characteristics of cost-effectiveness, disposability, miniaturization, wide potential window, and easy of electrode design. Compared with gold or platinum electrodes, surface modification is difficult because the carbon surface is chemically or physically stable. Oxygen plasma can easily produce carboxyl groups on the carbon surface, which act as scaffolds for covalent bonds. However, the effect of O2-plasma treatment on electrode performance remains to be investigated from an electrochemical perspective, and sensor performance can be improved by clarifying the surface conditions of plasma treated biosensors. In this research, we compared antibody modification by plasma treated and physical adsorption, using our original immunosensor based on gold nanoparticles (AuNPs). Consequently, the O2-plasma treatment produced carboxyl groups on the electrode surface, which changed the electrochemical properties owing to electrostatic interactions. It was also found that the surface became hydrophilic, inhibiting non-specific antigen adsorption. The sensitivity was 6.5 times higher than the Limit of Detection (LOD) using physical adsorption.