In this paper, capability of electrocentrifuge-spinning (ECS) method for generating highly aligned nanofiber was studied. First, the degree of nanofibers alignment (DNA) produced by ECS was compared with that of rotating drum (RD) method and ECS superiority was demonstrated. Then central composite design (CCD) and response surface methodology (RSM) was used for optimization of operating conditions. The critical factors selected for the examination were voltage, polymer concentration, collector diameter and spinneret rotational speed. To design the required experiments at the settings of independent parameters, RSM was applied. A total of 30 experiments were accomplished towards the construction of a quadratic model for target variable. Using this quadratic model, the influence of aforementioned variables was discussed on DNA. The best operating condition for attaining the maximum value of DNA was the applied voltage of 20.19kV, polymer concentration of 17.44wt%, collector diameter of 40.76cm, and rotational speed of 2680.10 rpm.