Preparation and Identification of Monoclonal Antibodies Against Der f1 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 f1 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 f1 allergen were obtained. The six McAbs all recognized recombinant Der f1 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 f1 allergen were prepared successfully.These antibodies may be used for the establishment of detection method of Der f1 allergen.