Preparation and identification of monoclonal antibody against Hendra virus N and G proteins
To prepare specific monoclonal antibodies against the Hendra virus(HeV),purified HeV-N and HeV-G proteins expressed in prokaryotic cells were used as antigens to immunize BALB/c mice.Spleen cells and S/P20 cells were fused using polyethylene glycol(PEG).Positive antibody hybridoma cells were screened using the indirect ELISA method.After three rounds of cloning,5 positive hybridoma cell lines were obtained,including three strains(named 2H6,3C2 and 3G6)targeting HeV-N protein and 2 strains(named 5B8 and 2A4)for HeV-G protein.Titers of the monoclonal antibodies were detected by ELISA method.The results showed that 2H6 and 2A4 were both ≥ 1∶512 000,3G6 and 5B8≥1∶256 000,and 3C2 ≥ 1∶128 000,respectively.West-ern blot and indirect immunofluorescence test results indicated that all 5 strains of monoclonal an-tibodies could recognize the corresponding antigens.The stability assay demonstrated that the cells were able to consistently secrete antibodies even after 15 generations,which illustrated that they had good stability.Monoclonal antibody subclass identification test revealed that 2H6,3C2 and 3G6 all belonged to the IgG2b subclass,whereas 5B8 and 2A4 were classified under the IgG1 subclass.The establishment of hybridoma cell lines secreting specific monoclonal antibodies against the N and G proteins of HeV laid a preliminary foundation for the development of new diagnostic meth-ods and reagents for HeV.
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