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Nowadays, the manual harvesting is the most common harvesting method of Lycium barbarum (L. barbarum), but the efficiency of the manual harvesting is low and the cost of that is high. In order to achieve the goal of high efficiency and low damage of the mechanical harvesting of L. barbarum, a portable vibrating and comb brushing harvester for L. barbarum based on the principle of vibrating and comb brushing was designed. Based on the separation principle of L. barbarum and the carpopodium, the mechanical model was adopted by single centralized mass method to establish the dynamic model of the harvester and the equations of the displacement, velocity and acceleration of contact point of ripe L. barbarum applied load by the harvester were obtained. In order to optimize the structure and working parameters of the harvester, the kinematics and dynamics simulation of the harvester was done through ADAMS. Results showed that the combination that the revolving speed of comb brush was 60～70r/min, the revolving speed of cam was 25～35r/min and the style of comb brush was Ⅱ, which was helpful to harvest L. barbarum. Three factors and three levels quadratic orthogonal revolving experiment was adopted to establish the mathematical model between the harvesting rate of ripe L. barbarum, harvesting rate of unripe L. barbarum, damage rate of ripe L. barbarum and revolving speed of comb brush, revolving speed of cam and style of comb brush. The influences of various factors on the harvesting rate of ripe L. barbarum, the harvesting rate of unripe L. barbarum and the damage rate of ripe L. barbarum were analyzed and the best combination was determined as the revolving speed of comb brush was 64.52r/min, the revolving speed of cam was 29.68r/min and the style of comb brush was Ⅱ. Field experiment results showed that the harvesting rate of ripe L. barbarum was 8912%, the harvesting rate of unripe L. barbarum was 5.87% and the damage rate of ripe L. barbarum was 6.24%. The research result provided a design basis for the later research of vibrating and comb brushing harvester for L. barbarum.