〔Abstract〕 Objective To develop a CD30-targeted CAR-T cell drug based on the multi-chain chimeric antigen receptor T cells(CAR-T) of the bridging protein DAP12, and to study thein vitroandin vivopreclinical efficacy of CD30 CAR-T on Hodgkin lymphoma tumor cells. Methods Through gene synthesis and molecular cloning techniques, a CAR plasmid targeting CD30 was designed and constructed, and the obtained lentivirus was packaged. The T cells were transfected with the lentivirus, where the multi-chain CAR-T targeting CD30 was the CD30-KIRS2/Dap12-BB group, the single-chain second-generation CAR-T was the CD30-41BBζ group, and the T cells without virus infection were the NTD group. The positive rate of CAR was detected by flow cytometry, the cytotoxicity of the cells was detected by lactate dehydrogenase(LDH) release assay, the secretion level of the cytokine interferon γ(IFN-γ) was detected by enzyme-linked immunosorbent assay(ELISA), and the antitumor activity of CD30 CAR-T in mice was further detected by a mouse xenograft tumor model. Results A comparison was made between the multi-chain CAR-T targeting CD30 and the single-chain second-generation CAR-T. It was found that the antitumor effect of the multi-chain CAR-T was similar to that of the single-chain CAR-T. However, it was worth noting that the IFN-γ secretion level of the multi-chain CAR-T was higher(P<0.001). More importantly, in the mouse tumor model experiment, the multi-chain CAR-T achieved complete tumor regression. Conclusion The multi-chain CAR-T targeting CD30 is superior to the traditional single-chain CAR-T in terms of antitumor activity.