The massive increase in the industrial revolution has now led to a decrease in the quality of the air, thereby increasing health problems. Air filtration has been recognized as an operational way to reduce particulate matter (PM) pollution for decades. Electrospun nanofibrous filters are regarded as an effective substitute over other air filter types due to their governable morphology with exceptional diameter/length ratio in a continuous method. In the current research, using electrospinning technique, 5 nanofibrous layers with varying concentrations were deposited over 3D printed polycarbonate (PC) support. A series of experiments were performed for electrospinning the PC nanofibers by altering various parameters such as nanofiber collection time, solvent ratio, deposition voltage, tip-to-collector distance, and polymer concentration. The filtration efficiency of the manufactured filters was evaluated, and its average fiber diameter, viscosity, conductivity, and pressure drop were measured. The nanofibrous layer exhibited a fiber diameter ranging from 0.19 ± 0.04 to 0.56 ± 0.14 μm. This nanofiber morphology enabled the filters to consistently achieve an efficient particle collection ability between 99.3 and 99.9%. Using the experimental results data, response surface plots were generated to visually represent the relationship between various factors and the desired characteristics of nanofiber media, including fiber diameter. The results show that all the nanofibers were successfully electrospun over 3D printed polycarbonate substrate and this filtering construction allowed for reaching high filtering efficiency (99.9%).