The paper presents a toll pricing methodology using a dynamic traffic assignment (DTA) system. This methodology relies on the DTA system’s capability to understand and predict traffic conditions, thus enhanced online calibration methodologies are applied to the DTA system, featuring a heuristic technique to calibrate supply parameters online. Improved offline calibration techniques are developed to apply toll pricing in a real network consisting of managed lanes and general purpose lanes. The online calibration methodologies are tested using real data from this network, and the results find the DTA system able to estimate and predict traffic flow and speed with satisfactory accuracy under congestion. Toll pricing is formulated as an optimization problem to maximize toll revenue, subject to network conditions and tolling regulations. Travelers are assumed to make route choice based on offline calibrated discrete choice models. Toll optimization is applied in a closed-loop evaluation framework where a microscopic simulator is used to mimic the real network. Online calibration of the DTA system is enabled to ensure good optimization performance. Toll optimization is tested under multiple experimental scenarios, and the methodology is found able to increase toll revenue compared with the condition when online calibration is not available. It should be noted that the toll rates and revenues presented in this paper are obtained in a simulation environment based on the calibration and optimization algorithms, and as the work is ongoing these results are far from being a recommendation to operators of managed lanes.