This study focuses on enhancing the mechanical strength of aquaporin (AQP)-based biomimetic membranes for seawater desalination. AQP incorporated vesicles were embedded into the selective layer of an optimized thin film composite (TFC) membrane. The resultant membrane, denoted as ASW, exhibited a stable water flux around 20 L.m(-2).h(-1) and 99% NaCl rejection at a constant pressure of 55 bar using 32,000 mg.L-1 NaCl solution as feed in reverse osmosis (RO) measurement. The robustness of the ASW membranes were evaluated. The water flux of ASW membrane was almost 100% enhanced compared with that of AQP-free control TFC membranes. The filtration performance of the ASW membrane was further evaluated by a seven-day desalination test using a real seawater secondary effluent collected from a desalination plant in Singapore as feed. To our best knowledge, our study is the first report on the AQP-incorporated RO membrane applied for seawater desalination. A commercial SW30HR membrane was tested in parallel for comparison. The robust ASW membrane exhibited a nearly 80% higher water flux in comparison to the SW30HR membrane with a comparable overall solute rejection, suggesting the advantage and feasibility of Aquaporin based biomimetic membranes for seawater desalination.