Knowledge of exchange flow and its variations is important, because it determines the distribution of salt, nutrient, pollutant, and suspended sediment in estuaries. Although exchange flow has been studied extensively, their variations over the spring-neap tidal cycle remain unclear. Many studies have indicated that the exchange flow is weaker during spring tides than neap tides, but others have reported that it might be stronger during spring tides. Therefore, we investigated the spring-neap variation of an exchange flow and its cause based on intensive observational data and an analytical model applied to the Sumjin River estuary (SRE), Korea. The observations revealed that exchange flow increased during the spring tide but decreased during the neap tide. The horizontal salinity gradient increased about seven times more during spring tide than during neap tide in the middle of the estuary. The analytical model implied that the exchange flow in the SRE is mainly driven by the horizontal salinity gradient. The large horizontal salinity gradient and tidal current amplitude ratio in the SRE between the spring and neap tides suggests that the horizontal pressure gradient driven by the salinity gradient overwhelms vertical mixing in driving exchange flow in the SRE.