A common challenge among internal combustion engine (ICE) manufacturers is shortening the development time while facing requirements and specifications that are becoming more complex and border in scope. Virtual simulation and calibration are effective instruments in the face of these demands. This article presents the development of zero-dimensional (0D)—real-time engine and exhaust after-treatment system (EAS) models and their deployment on a Virtual test bench (VTB). The models are created using a series of measurements acquired in a real test bench, carefully performed in view of ensuring the highest reliability of the models themselves. A zero-dimensional approach was chosen to guarantee that models could be run in real-time and interfaced to the real engine Electronic Control Unit (ECU). Being physically based models, they react to changes in the ECU calibration parameters. Once the models are validated, they are then integrated into a Simulink® based architecture with all the Inputs/Outputs connections to the ECU. This Simulink® model is then deployed on a Hardware in the Loop (HiL) machine for ECU testing and calibration. The results for engine and EAS performance and emissions align with both steady-state and transient measurements. Finally, two different applications of the HiL system are presented to explain the opportunities and advantages of this tool integrated within the standard engine development. Examples cited refer to altitude calibration activities and soot loading investigation on vehicle duty cycles. The cases described in this work are part of the actual development of one of the latest engines developed by Kohler Engines: the KDI 1903 TCR Stage V. The application of this methodology reveals a great potential for engine development and may become an essential tool for calibration engineers.