The Dual Stage Pressure Retarded Osmosis technique is considered for power generation. The influence of feed flow rates, hydraulic pressure, and pressure drop on mass transfer and solute diffusion in a full-scale membrane model was investigated for the first time to maximize power generation. Dead Sea-seawater, Dead Sea-reverse osmosis brine, reverse osmosis brine-wastewater, and seawater-wastewater salinity gradient resources were investigated for power generation. Results revealed a 71.07% increase in the specific power generation due to the dual-stage pressure retarded osmosis process optimization using Dead Sea-seawater salinity gradient resources. The increase in the specific power generation due to the dual-stage pressure retarded osmosis optimization was 108.8%, 63.18%, and 133.54%, respectively, for Dead Sea-reverse osmosis brine, reverse osmosis brine-wastewater, and seawater-wastewater salinity gradient resources. At optimum operating conditions, using the dual-stage pressure retarded osmosis process as an alternative to the single pressure retarded osmosis process achieved up to a 22% increase in the energy output. Interestingly, the hydraulic pressure at optimum operating conditions was slightly higher than the average osmotic pressure gradients in the dual-stage pressure retarded osmosis process. The study also revealed that power generation in the dual-stage pressure retarded osmosis process operating at constant mass transfer and solute resistivity parameters was overestimated by 2.8%.