Résumé
Three BALB/c mice were immunized four times with alpha-momorcharins (alpha-MMC). Using polyethylene glycol (PEG) method, the immunized splenocytes were fused with SP2/0 cells. One strain of hybridoma cells was obtained which secrete antibodies against alpha-MMC. To get ascites, the hybridoma cells were injected into the abdominal cavity of mice. The antibodies were purified from ascites. Indirect enzyme linked immunosorbent assay (ELISA) and Western blot assay were applied to determine the specifity of the monoclonal antibody (McAb). The results showed that the McAb was specific to alpha-MMC without detectable cross-activity with MAP30. The McAb provided detecting method for further research of the structure and function of alpha-MMC.
Sujets)
Animaux , Femelle , Souris , Anticorps monoclonaux , Allergie et immunologie , Technique de Western , Test ELISA , Hybridomes , Métabolisme , Souris de lignée BALB C , Protéines inactivant les ribosomes , Allergie et immunologieRésumé
Both IFN-omega and IFN-alpha belong to type I interferon and have antiviral, antiproliferative, immunomodulatory activities, but their bioactivities are usually different. FeIFN-omega gene was amplified by PCR. FeIFN-alpha gene was synthesized based on the published sequences of GenBank. Then the two types of feline interferon genes were subcloned into the pET-His vector, and expressed in Escherichia coli Rosetta (DE3). Recombinant interferons were purified by affinity chromatography with immobilized nickel chelating NTA (Ni-NTA) and their antiviral activity was estimated according to the ability of IFNs to inhibit the cytopathic effects (CPE) of virus on cells. Results showed that the antiviral activities against various viruses of FeIFN-omega were higher than those of FeIFN-alpha. Against H9N2 subtype avian influenza virus (AIV) and canine distemper virus (CDV), the antiviral activities of FeIFN-omega were 160 folds and 4 folds higher than those of FeIFN-alpha.