This study synthesized collagen-chitosan/PVA nanofiber composites using the electrospinning method. Characterizations included Fourier transform infrared spectroscopy (FTIR) and surface morphology using scanning electron microscopy (SEM). Investigations were carried out on tensile strength, degradation rate, and antibacterial test. It was found that the functional groups C–H, –OH, C–O, C–N, and N–H were suitable for PVA, collagen, and chitosan materials. The SEM showed that increasing the PVA composition caused a change in fiber diameter ranging from 34.64 to 71.63 nm. The tensile strength results show that the smallest nanofiber diameter has the highest ultimate strength value of 5.6 ± 0.4 MPa. In addition, it was found that the rate of degradation was in the range of 44%–76%. The antibacterial activity test was carried out using two types of bacteria, i.e., a gram-positive bacteria, S. aureus, and a gram-negative bacteria, E. coli. The test results showed that the collagen-chitosan/PVA nanofiber composite had a diameter of antibacterial inhibition for E. coli and S. aureus bacteria, respectively.