Hemodialysis is one of the most commonly used treatments for patients suffering from irrecoverable kidney damage. In our present work, we investigate poly(ether imide) (PEI) mixed matrix membranes (MMMs) as a potential candidate for hemodialysis applications due to their efficient clearance and high biocompatibility. Graphene oxide (GO) was synthesized by the modified Hummers' method and was then confirmed by X-ray diffraction spectroscopy, Fourier transform infrared spectroscopy, Raman spectroscopy, and high-resolution transmission electron microscopy. The GO-polyvinylpyrrolidone nanocomposite incorporated PEI MMMs were fabricated by a semiautomatic casting unit using the nonsolvent induced phase separation technique. The effect of the nanocomposite loading ratio was evaluated by water content, ultrafiltration rate, and porosity, which were all found to increase as the nanocomposite content increased. Cross-sectional and top surface morphology was visualized using scanning electron microscopy and atomic force microscopy. The hydrophilicity of these membranes was in consonance with contact angle values. These MMMs demonstrated an increase in biocompatibility: reduced protein adsorption, suppressed platelet adhesion, and lower complement activation. Furthermore, the prolonged blood clotting time is an indication of the heparin mimic anticoagulant properties of these membranes. The cytocompatibility results by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-tetrazolium bromide assay and live cell/dead cell staining indicated that there was an increase in cell viability. The membranes with 0.1 wt % GO showed an excellent clearance of the model uremic toxins, namely urea, vitamin B-12, and cytochrome-c in vitro. The diffusive permeability of these membranes could be comparable to the existing commercial hemodialysis membranes. Thus, it can be concluded that these membranes containing a composite of both functional nanosheets and bioactive polymers have a tremendous potential to be utilized commercially in hemodialysis modules if shown successful in further in vivo studies with an animal model.