An analysis of antigenic polypeptides of dengue viruses and their specificity with western blotting

Abstract

Dengue virus(DV) samples which harvested after having been either propagated in C₆/36 cells or passaged by murine intracerebral inoculation were investigated by Western bloting. Two different denaturing methods with the same solution were selected before electrophoresis, one at 60 °C for 20 min; the other at 100°C for 5 min. After the samples were heated at 60°C for 20 min, 3 bands of sizes 46 ku, 70 ku and 98 ku were positively recognized by rabbit immunized sera. The peptide of 98 ku was DV group specific; While that of 46 ku could also react with the sera against both heterologous DV serotype and Japanese B encephalitis virus (JEV), After the samples were heated at 100 °C for 5 min, 3 peptides in size of 20 ku, 46 ku and 57 ku were recognized by the rabbit sera against both homologous and heterologous DV serotypes, while the peptide of 57 ku could also be recognized by anti-JEV serum. There was no significant difference in the results of Western blot between the DV antigens harvested from cellular culture and those from intra-cerebral inoculation.

Keywords

dengue viruses / antigenic polypeptides / western blot / antigenic epitopes

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