The successful application of bioventing systems for in situ remediation of petroleum hydrocarbon-contaminated sites depends on many factors including: moisture content, temperature, number and location of injection and extraction wells, as well as corresponding airflow rates. The importance of well locations and air flow rates has been explored in this study and their optimal values were obtained via the fixed well method. The design problem was formulated to maximize total contaminant mass removal within a given time frame subject to physical and budgetary constraints. The technique developed herein was applied to a field case study. Results indicate that optimal well locations are found along the mass centerline(s) of the contaminant plume. Furthermore, well location optimization was proven to be more important than pumping rate optimization.