Point-of-Care (POC) devices used for medical applications such as disease detection, monitoring, and treatment, require certain levels of accuracy, efficiency, reliability, and accessibility. In order to meet these requirements, this study presents the development of a flexible electrochemical biosensor with a novel design and fabrication technique. This biosensor utilizes cost-effective and available materials, while incorporating surface modification techniques using graphene and gold nanoparticles to enhance sensitivity. The improved performance of the developed device is demonstrated through a comprehensive set of optical and electrochemical experiments, showing enhanced sensitivity, durability, repeatability, and limit-of-detection for electrochemical enzyme-linked immunosorbent assay (e-ELISA) targets. The findings of this study highlight the potential of our biosensor to be utilized in wearable applications, thereby improving patient outcomes, and reducing healthcare costs.