本研究制备并鉴定了鼠抗粉尘螨变应原Der f 5的单克隆抗体(monoclonal antibody，McAb)。将质粒pET28a(+)-Der f 5转化ArcticExpress(DE3)感受态细胞，取单个菌落接种培养，加入IPTG诱导表达，经纯化获得Der f 5重组蛋白。利用Der f 5重组蛋白为免疫原，免疫BALB/c小鼠，取免疫小鼠脾脏细胞与其骨髓瘤细胞融合。通过间接ELISA法筛选特异性分泌抗体的杂交瘤细胞。用杂交瘤细胞株诱生小鼠腹水，应用蛋白A亲和层析法进行抗体纯化。通过间接ELISA、Western blotting鉴定该单克隆抗体的特性和交叉性。最终获得3株稳定分泌鼠抗粉尘螨变应原Der f 5的高效价单克隆抗体。研究分析表明该3株单抗均能识别重组Der f 5蛋白和天然的粉尘螨提取物。本实验成功制备了3株鼠抗粉尘螨变应原Der f 5的单克隆抗体，为建立粉尘螨主要变应原Der f 5检测及纯化方法奠定了基础。

To prepare monoclonal antibodies against allergen Der f 5 from Dermatophagoides farinae and to characterize their properties, recombinant plasmid pET28a(+)-Der f 5 was transformed into ArcticExpress (DE3) competent cells. The recombinant colonies were inoculated with IPTG and purified to obtain Der f 5 recombinant protein. BALB/c mice were immunized with the purified recombinant Der f 5 allergen, and the splenocytes of the immunized mice were fused with NS-1 myeloma cells by hybridoma technique. Indirect ELISA was used for screening the hybridoma cell lines which could secrete antibodies against Der f 5. The McAbs against Der f 5 allergen were purified using affinity chromatography on immobilized protein A to identify their specificity, titers and cross-reactivity by ELISA and Western blotting. Three hybridoma cell lines secreting the McAbs against Der f 5 allergen were obtained, which had good ascites titers. The three McAbs against Der f 5 allergen all recognized recombinant Der f 5 allergen and native extract from Dermatophagoides farinae. Therefore, three monoclonal antibodies against Der f 5 allergen were prepared successfully, which will facilitate establishing detection and purification method of Der f 5 allergen.