Preparation of monoclonal antibodies against recombinant Der f 7 allergen

VernacularTitle:粉尘螨变应原第7组分(Der f 7)重组蛋白单克隆抗体的制备
Author: Jinxia SUN ; Lili YU ; Feixiang TENG ; Li YANG ; Yubao CUI
Keywords: Der f 7; Monoclonal antibody; Dust mites; Allergen
From: Chinese Journal of Immunology 2017;33(7):1039-1042
CountryChina
Language:Chinese
Abstract: Objective:To clone and express the Der f 7 recombinant protein from Dermatophagoides farinae and prepare the Der f 7 monoclonal antibody.Methods: The Der f 7 recombinant protein was expressed in prokaryotic expression system of pET28a(+)-Der f 7.BALB /c mice were immunized with the recombinant protein.Myeloma cells and spleen cells were fused,and hybridoma cells were screened by ELISA.Hybridoma cells were injected into the mice abdominal cavity to obtain ascites.It was purified by protein A agarose medium ascites,and then to dentified the titer and purity of the antibody.The specificity of the antibody was identified by Western blot.Results: Three hybridoma cells which stably secret recombinant Der f 7 monoclonal antibody were obtained.The monoclonal antibody had high purity,the titer was higher than 1∶243 000.Western blot showed that Der f 7 recombinant protein could be recognized well.Conclusion: We successfully obtained Der f 7 monoclonal antibody,which can be used for the quantification and localization of Der f 7 allergen and the diagnosis and treatment of allergic diseases.