A new fouling-resistant mixed-matrix nanofiltration membrane based on a covalently cross-linked Ti3C2TX (MXene)/cellulose acetate (MXene@CA) composite was fabricated by phase inversion followed by formaldehyde cross-linking. The physicochemical properties of the prepared MXene@CA composite membranes were studied by field emission scanning electron microscopy (FESEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD) and water contact angle techniques. The performance of the prepared membranes was evaluated with respect to the water flux, bacterial growth inhibition and rejection properties. The 10%MXene@CA (10:90 wt % of MXene:CA) composite membrane shows high pure water flux of $\sim$256.85 L m−2 h−1 bar−1, 123.28% water uptake and 69.7% porosity. The 10%MXene@CA membrane, exhibited more than 92% and 98% rejection of rhodamine B (RhB) and methyl green (MG), respectively. Furthermore, 10%MXene@CA membrane exhibited more than 98% and 96% growth inhibition for E. coli and B. subtilis, respectively. Also, the optimal membrane showed a significantly improved hydrophilicity (water contact angle = 60.8°), which has favored good antifouling properties. The reported nanofiltration membrane, especially 10%MXene@CA, can be suggested for water purification and biomedical applications.