The capability of misalignment tolerance is an important requirement for the capture of the cooperative target, which can offset the position and orientation errors of the cooperative target that is caused by the space manipulators. A reusable lock and separation device with a certain misalignment tolerance is presented and designed according to the capture way of the cooperative target of on orbit service platform, which can lock and separate the cooperative target under the assistance of the space manipulator. The D-H matrix between the arbitrary point of the cooperative target and the coordinate system of reusable lock and separation device is established. In addition, the mathematical model of the trajectory of key points of the ball head is completed. In order to verify the feasibility of the device design, the misalignment tolerance is analyzed by the presented way of the projection of key points. The dynamics simulation of the misalignment tolerance performance is carried out through ADAMS software. Besides, the experiment of the prototype is carried out. The reusable lock and separation device of misalignment tolerance is validated through the simulation and experimental results.