A remote-control tether-less isolation tool is a mechanical device that is normally used in pipelines to block the flow at a given position by transforming a blocking module. In this study, the interactions between the fluid and the plug module of the isolation tool were investigated. Simulations of the plug process and particle image velocimetry measurements were performed to study the flow characteristics. Numerical solutions for the continuity, momentum, and energy equations were obtained by using commercial software based on finite-volume techniques. Box–Behnken design was applied, and response surface methodology (RSM)-based CFD simulation analysis was conducted. The dynamic model in the plug process was built by RSM and used to evaluate the influences of the main mechanical parameters on the pressure during the plug process. The diameter of the isolation tool and the diameter of the plug module have strong influences on the process, and the length of the isolation tool has only a little influence on the plug process.