In order to find the optimal combination of the structure and working parameters of comb type cotton picker harvesting device, the experiments were conducted. A central composite rotatable orthogonal experimental design of response surface methodology was employed to find the optimum combination effect on working properties. The three parameters, including working speed, comb space and vibration frequency of comb, were selected as input variables. Picking rate and loss rate were selected as response functions. Experimental results indicated that the most optimum combination region was: working speed of 0.45 m/s, comb space of 51 mm, and comb vibration frequency of 2.65 Hz, the picking rate was 96.5%, the loss rate was 2.8%.