In the present study, novel silk fibroin (SF)/polyacrylonitrile (PAN) double-layer nanofilters defined by a variable composition of polyaniline (PANI)/TiO2 nanoparticles were prepared using an electrospinning technique, and their application for dye removal was investigated. Scanning electron microscopy (SEM) showed that fibrous mats characterized by nanoscale diameters (70-120 nm) and random ultrafine fiber orientations without beads were successfully obtained. Tensile test results revealed that the strength and Young's modulus were markedly enhanced in the hybrid double-layer SF-PAN membranes relative to single-layer polymers. In addition, the incorporation of PANI/TiO2 nanoparticles within the hybrid samples resulted in a notable increase in strength and modulus. The adsorption-assisted nanofiltration performance of the membranes in removing anionic reactive black (HFGR) dye from aqueous solution was investigated using a dead-end stirred cell filtration device. The effect of several variables, including PANI/TiO2 nanoparticle content, pH and dye concentration, were evaluated. Hybrid nanofilters containing 7.5 wt% PANI/TiO2 nanoparticles demonstrated excellent dye removal efficiency of up to similar to 92% in acidic solution.