In view of the characteristics of the poor driving comfort for beam bridges, to explore the passing vehicle ride comfort in the process of driving on the highway bridge, a vehicle-bridge coupling vibration differential equation was established with a beam bridge as the analysis object, based on the theory of vehicle bridge coupling vibration. The seat acceleration of the vehicle was calculated by using the self-programmed vehicle bridge coupling MATLAB program, and the ride comfort of the driver and passenger was evaluated by the international standard ISO2631-1 root mean square acceleration value evaluation method. The influences of vehicle type, seat position, vehicle weight, bridge surface roughness, speed and other factors on the ride comfort of the vehicle were analyzed. The results show that the driving comfort of cars is better than that of trucks and buses for different vehicle types. For a two-axle bus with multiple rows of seats, the ride comfort of the front seat is better than that of the back seat in different riding positions, and the farther the seat is from the vehicle centroid, the more comfortable the ride is. If different passengers are in the same row, the comfort of standing passengers is worse than that of seat passengers. Vehicle ride comfort is very sensitive to bridge deck road condition, and it decreases rapidly with the deterioration of bridge deck road condition. For different vehicle weight, vehicle ride comfort increases with the increase of vehicle weight. Vehicle speed has a certain influence on ride comfort with less effect. It is suggested that beam bridge vehicle speed limit measures be taken in order to improve the ride comfort.