In a 3-year research project, a new approach to forecast biological N2O formation and emission at high-strength reject water treatment has been developed (ASM3/1_N2OISAH). It was calibrated by extensive batch-tests and finally evaluated by long-term measurement campaigns realized at three wastewater treatment plants (WWTPs) with different process configurations for nitrogen removal of reject water. To enable a model application with common full-scale data, the nitritation-connected supplementary processes that are responsible for N2O formation are not depicted in the model. Instead, within the new model approach the N2O formation is linked to the NH4-N oxidation rate by defining specific formation factors [N2O-Nform/NH4-Nox], depending on the concentrations of NO2 and O2 as well as the NH4 load. A comparison between the measured and the modeled N2O concentrations in the liquid and gas phase at the full-scale treatment plants prove the ability of the proposed modelling approach to represent the observed trends of N2O formation, emission and reduction using the standard parameter set of kinetics and formation factors. Thus, enabling a reliable estimation of the N2O emissions for different operational conditions. The measurements indicate that a formation of N2O by AOB cannot completely be avoided. However, a considerable reduction of the formed N2O was observed in an anoxic environment. Applying the model, operational settings and mitigation strategies can now be identified without extensive measurement campaigns. For further enhancement of the model, first results for kinetics of N2O reduction kinetics by denitrification processes were determined in laboratory-scale batch tests