Performance of the nanofibrous filter media is determined by fibre diameter and pore size distribution of the membranes. In this study, an attempt is made to fabricate a nanofibrous multi-scale filter media by simply varying the layer-by-layer-assisted stacking structure and composition of individual fibres within the single layers. These structural controls of the filter membrane helped to form a skeletal frame-worked membrane with controlled filtration efficiency and pressure drop aerosol particle filtration application. Altering the right electrospinning parameters, layering structure of the membrane resulted in changes in average fibre diameter and pore size distribution, especially in the middle layer of media, while a gradient in pore size and its distribution was created. The results of measurements of pressure drop and filtration efficiency indicated that by changing the gradient in pore size and its distribution in the middle membrane, keeping other layers constant, the pressure drop and filtration efficiency can be effectively tuned. The results suggested that this developed filter media could be used as promising materials for a variety of potential applications in high efficiency particulate air filters with tunable characteristics.