Development of point-of-care biosensors continues to gain popularity due to the demand of improving the cost performance in today’s health care. As cardiovascular disease induced death remains on the top 3 death causes for most Asian countries, this paper is to present a high-sensitivity point-of-care biosensor for the detection of cardiovascular disease biomarkers. To meet the point-of-care biosensors requirements, which include characteristics such as small size, low cost, and ease of operation, we adopted electrochemical methods as the basis of detection. The 4-aminothiophenol was adopted as the bio-linkers to facilitate the antibody-antigen interaction. A more stable three-electrode configuration was miniaturized and laid out onto a biochip. A microfluidics subsystem based on opto-piezoelectronic technology was also integrated to create the microfluidic biochip system. To improve the detection sensitivity associated with the reduction in biochip size, electrochemistry simulation was used to investigate several potentially effective means. We found that the electric field on the edge near working electrode and counter electrode was higher, which was verified by using atomic force microscopy to measure the surface potential. With the successful verification, we explored the configuration, i.e., lengthened the edge of working electrode and counter electrode without changing the areas of working electrode and counter electrode and the gap between these two electrodes, so as to evaluate the possibility of improving the measurement efficiency in our newly developed biochips. Detailed design, simulation and experimental results, improved design identified, etc. were all presented in detail.