Peptide based polyclonal antibody production against bovine rotavirus non structure protein4 (NSP4)

The rotavirus nonstructural protein 4 (NSP4), is a multi functional protein that play key role in both viral morphogenesis and cytopathic effect associated with cell death. However, the complete biological effect of NSP4 remains to be clarified. Since to obtain further knowledge about this protein there is a need for recognizing antibody and there is no commercial antibody against this protein, this study was designed to produce polyclonal antibodies against a synthetic immunogenic peptide of RF rotavirus NSP4 protein in order to be used as diagnostic and research tool. In the present study a peptide sequences corresponding to NSP4 114-135 conjugated to Bovine Serum Albumin (BSA) by glutaraldehyde through a single-step coupling protocol. The conjugated peptides were extensively dialyzed and injected to New Zealand white rabbits. The NSP4 114-135 peptide-specific antiserum was confirmed by both IF and Western blot analysis. The result indicated successful production of polyclonal antibody raised against native NSP4 protein. In conclusion, since NSP4 is a toxic protein for the cells, it is impracticable to express full length of NSP4 protein in expression systems. Therefore, the introducing immunogenic peptide in appropriate animal may be the best approach to produce its specific antibody.