The invention provides new application of rabbit fetal fibroblast, and particularly relates to a method for preparing a ZEN (Zearalenone) monoclonal antibody by utilizing the rabbit fetal fibroblast.The method is characterized in that mouse hybridoma is cultured by using rabbit fetal fibroblast as feeder cells of murine hybridoma, the murine feeder cells can be completely replaced, growth of monoclonal hybridoma can be promoted, and a monoclonal antibody with high valence, good stability and strong specificity can be generated; the hybridoma bred by different animal cells as the feeder cellsis used for preparing an antibody, and very high valence and very high specificity are obtained; meanwhile, the rabbit fetal fibroblast can be revived by many times for use after being cryopreserved,animal tissue isolating culture is not required to be carried out every time. The method disclosed by the invention is simple, the cost is low, the monoclonal antibody which is used for fast, sensitive, economic and efficient ZEN immunological detection can be prepared, and a wide application prospect and huge market value are obtained.