Monoclonal antibodies production specific to vitellin and vitellogenin of giant tiger prawn Penaeus monodon

Abstract

Summary Monoclonal antibodies specific to Penaeus monodon vitellin and vitellogenin were produced using crude ovarian extract from gravid P. monodon ovaries. After immunization and fusion of mouse spleen cells with P3X myeloma, hybridomas were selected by indirect immunoperoxidase ELISA against P. monodon ovarian extract, followed by dot-blotting against native and denatured proteins from ovarian extract, female haemolymph, and male haemolymph. Four hybridoma clones producing antibodies (PMV-11, 15, 22 and 64) were identified. They bound with ovarian extract proteins and with female haemolymph, but not with male haemolymph. One antibody (PMV-64) bound with both native and denatured proteins. Using Western Blot analysis of ovarian extract separated by PAGE, all four monoclonal antibodies bound with the same lipoglycoprotein band. Using Western Blot analysis of proteins separated by SDS-PAGE, PMV-64 antibody bound with Mw 80 and 83 kD proteins in ovarian extract, and with Mw 83 and 200 kD proteins in female haemolymph. All four monoclonal antibodies belong to the IgG1 subclass.