Abstract: Positioning of mobile robots basically calculated using odometry information. Odometry from the wheel’s encoder is mostly used for simple and inexpensive implementation for determining the relative localization of a mobile robot. This paper deals with the estimation of better relative localization of a two wheeled differential drive robot by means of odometry by considering the influence of parameters namely payload, speed, diameter of wheel and thickness of wheel. Experiments have been conducted based on central composite rotatable design matrix. A mathematical model has been developed for the robot using Response Surface Methodology (RSM) with the help of MINITAB software. An optimum relative positioning was obtained by using Excel Solver.