In a previously published study, to optimize the vibrating and comb-brushing harvesting, the main factors and their parameter values were obtained based on the FEM and RSM. However, the study was based on the extensive cultivation mode which need to be improve. To realize the mechanization of the harvesting of Lycium barbarum L., as well as to face the standardized hedge cultivation mode, a vibrating and comb-brushing harvester machine was designed, which was primarily composed of an execution system, a motion system, and a control system. The mathematical model between the harvest effect index and the operation parameters was established based on response surface methodology (RSM). The effects of various parameters on the harvest index were analyzed, and the best parameter combination was determined: a vibration frequency of 38.73 Hz, a brush speed of 14.21 mm/s, and an insertion depth of 26.07 mm. The field experiment showed that the harvesting rate of ripe fruit was 83.65%, the harvesting rate of unripe fruit was 7.22%, the damage rate of the ripe fruit was 11.49%, and the comprehensive picking index was 87.85. The findings provided a reference for the development of L. barbarum harvesting mechanization in a standardized hedge cultivation mode.