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The current study reports the synthesis of graphite-based poly (acrylonitrile-co-acrylic acid) composite (GPC) sheets via sol-gel/casting method. The effect of different amounts of graphite (0.25 to 40 wt%) on various characteristics of the prepared GPC was studied through scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), and differential thermal gravimetric analysis (TGA/DTG). The XRD and FTIR analyses confirmed the existence of characteristic peaks of graphite while SEM micrographs showed its uniform distribution in polymer matrix. TGA curves revealed enhancement of thermal stability of GPC with increasing graphite content. The swelling ratio and water holding capacity analyses revealed a reduced hydrophilic behavior of GPC with increasing graphite content. The fabricated GPC demonstrated impressive antimicrobial properties against Escherichia coli. The structural, physico-chemical, thermal, and biological features of as-prepared GPC show their potential biomedical applications.