抗粉尘螨主要变应原Der f1单克隆抗体的制备与鉴定

目的制备和鉴定鼠抗粉尘螨主要变应原Derf1的单克隆抗体（monoclonal antibody，McAb）。方法利用重组Derf1蛋白为免疫原，免疫BALB／c小鼠，取免疫小鼠脾细胞与小鼠骨髓瘤NS-1细胞融合。通过间接ELISA法筛选特异性分泌的杂交瘤细胞。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。采用Ig类与亚类鉴定试剂盒鉴定该单克隆抗体的Ig亚型；通过间接ELISA、Western Blotting鉴定该单克隆抗体的特性和交叉性。结果获得6株可稳定分泌鼠抗粉尘螨主要变应原Derf1的单克隆抗体，其Ig亚型均为IgG1，且6株单抗均具有良好的效价。ELISA和Western—blotting分析表明，该6株单抗均能识别重组Derf1蛋白和天然的粉尘螨提取物。其中，除了484不识别屋尘螨提取物，其他均能识别。结论成功制备了6株鼠抗粉尘螨主要变应原Derf1的单克隆抗体，为建立粉尘螨主要变应原Derf1检测及纯化方法奠定了基础。

Preparation and Identification of Monoclonal Antibodies Against Der fl Allergen

Objective To prepare monoclonal antibodies against Der f 1 allergen from Dermatophagoides farinae and to characterize their properties. Methods BALB/c mice were immunized with purified recombinant Der f1 allergen protein, and the splenocytes of the immunized mice were fused with NS-1 myeloma cells by hybridoma technique. The McAbs against Der fl allergen were purified using affinity chromatography on immobilized Protein A and identified by their specificity, subtype, titers and cross-reactvity via ELISA and Western-blotting. Results Six hybridoma cell lines secreting IgG1 subtype McAbs against Der fl allergen were obtained. The six McAbs all recognized recombinant Der fl allergen and the native proteins extracted from Dermatophagoides farinae.With the exception of 4B4 McAb, the other five McAbs also recognized the native proteins extracted from Dermatophagoides pteronyssinus. Conclusion Six monoclonal antibodies against Der fl allergen were prepared successfully.These antibodies may be used for the establishment of detection method of Der fl allergen.