Biofilm models are valuable tools for process engineers to simulate biological wastewater treatment. In order to enhance the use of biofilm models implemented in contemporary simulation software, model calibration is both necessary and helpful. The aim of this work was to develop a calibration protocol of the particulate biofilm model with a help of the sensitivity analysis of the most important parameters in the biofilm model implemented in BioWin (R) and verify the predictability of the calibration protocol. A case study of a circulating fluidized bed bioreactor (CFBBR) system used for biological nutrient removal (BNR) with a fluidized bed respirometric study of the biofilm stoichiometry and kinetics was used to verify and validate the proposed calibration protocol. Applying the five stages of the biofilm calibration procedures enhanced the applicability of BioWin (R), which was capable of predicting most of the performance parameters with an average percentage error (APE) of 0-20%. (C) 2012 Elsevier Ltd. All rights reserved.