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.