Diversifying energy resources and reducing greenhouse gas emissions are key priorities in the forthcoming years for the automotive industry. Currently, among the different solutions, sustainable biofuels are considered as one of the most attractive answer to these issues.This paper deals with the vehicle application of an innovative diesel fuel formulation using Ethanol to tackle these future challenges. The main goal is to better understand the impact of using biofuel blends on engine behavior, reliability and pollutants emissions. This alternative oxygenated fuel reduces dramatically particulate matter (PM) emissions; this paves the way to improve the NOx/PM/CO₂ trade-off. Another major interest is to avoid adding a particulate filter in the exhaust line and to avoid modifying powertrain and vehicle hardware and therefore to minimize the overall cost to fulfill upcoming emission regulations.After a first part devoted to a description of the fuel properties, the paper focuses on some combustion analysis pointing out the effects of the new formulation. Then the fuel/engine compatibility and its optimization on a production turbocharged diesel multi-cylinder engine with high pressure exhaust gas recirculation are investigated. Owing to advanced calibration methods such as design of experiments, new maps are provided to fully take advantage of the ethanol/diesel blend properties and to enhance the engine behavior. It leads to a simultaneous decrease of nitrogen oxides and particulate matter emissions with a potential CO₂ emissions reduction and combustion noise decrease. The main drawbacks could be an increase of unburned hydrocarbons and carbon monoxide emissions.Furthermore, after having successfully validated the new settings on the test bench, some vehicle results on a chassis dynamometer are presented on the New European Driving Cycle (NEDC) to illustrate the effectiveness of this innovative fuel formulation and the calibration adaptation. Finally the recalibrated vehicle, integrating only Euro 4-compliant technologies (not equipped with a diesel particulate filter device), with this new fuel satisfies potentially Euro 5 emissions standards without sacrificing driveability and service life.