Many modeling and calibration methods have been developed to analyze and design the biological wastewater treatment process. For the systematic use of activated sludge model (ASM) in a real treatment process, a most important step in this usage is a calibration which can find a key parameter set of ASM, which depends on the microorganism communities and the process conditions of the plants. In this paper, a standardized calibration protocol of the ASM model is developed. First, a weighted effluent quality index(WEQI) is suggested far a calibration protocol. Second, the most sensitive parameter set is determined by a sensitive analysis based on WEQI and then a parameter optimization method are used for a systematic calibration of key parameters. The proposed method is applied to a calibration problems of the single carbon removal process. The results of the sensitivity analysis and parameter estimation based on a WEQI shows a quite reasonable parameter set and precisely estimated parameters, which can improve the quality and the efficiency of the modeling and the prediction of ASM model. Moreover, it can be used for a calibration scheme of other biological processes, such as sequence batch reactor, anaerobic digestion process with a dedicated methodology.