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In this report a novel antibacterial wound dressing was prepared and then characterized for required testing. We loaded silver sulfadiazine (AgSD) for the first time by electrospinning. AgSD was added in zein (0.3%, 0.4%, 0.5%, and 0.6% by weight) and was electrospun to fabricate nanofiber mats for wound dressings. Nanofiber mats were characterized by Fourier transform infrared spectroscopy (FTIR) to check if there was any chemical reaction between AgSD and zein. Morphological properties were analyzed by Scanning Electron Microscopy (SEM), which showed uniform nanofibers without any bead formation. The diameter of the nanofibers gradually decreased with an increase in the amount of AgSD, which can be associated with strong physical bonding between zein and AgSD. Thermal properties of nanofiber mats were analyzed by Thermogravimetric Analysis (TGA). X-Ray Diffraction (XRD) further demonstrated the crystalline structure of the nanofiber mats, and X-ray Photoelectron spectroscopy (XPS) was performed to confirm Ag and S contents in the prepared wound dressings. In order to investigate antibacterial properties, a disc diffusion method was employed. <i>Bacillus</i> and <i>E. coli</i> bacteria strains were used as Gram-positive and Gram-negative strains respectively. The antibacterial effectiveness of AgSD released from zein nanofibers was determined from the zone inhibition of the bacteria. The antibacterial activity of zein nanofibers loaded with drug was observed with both strains of bacteria in comparison to a control. Excellent antibacterial efficacy was attributed to the sample with 0.6% AgSD. Excellent release properties were also associated with the sample with 0.6% AgSD in zein nanofibers. Keeping in mind the abovementioned characteristics, prepared nanofiber mats would be effective for application in wound dressings.