Engineered Recombinant NDV-Fusion Protein and Its Polyclonal Antibodies Production

Abstract

Antibodies have important role in biological research and diagnosis. Polyclonal antibodies can be produced against more than one epitope of an antigen. Newcastle disease (ND) is one of the most devastating diseases that considerably effects the global poultry industry. Newcastle disease virus (NDV) possess Fusion (F) protein for the attachment and pathogenicity. In the present study TA plasmid having F gene was restricted by EcoR1 and it was ligated into pET30a expression vector. The F gene ligated pET30a was transformed in BL21DE3 expression strain. After the expression of NDV F protein, it was verified by the appearance of 67 kDa band on SDS-PAGE gel and western blot. The F protein was partially purified by the column of 70 kDa cut. The recombinant F protein was used for the generation of polyclonal antibodies. Two groups (experimental group and negative control group) of 3 months old rabbits were used. Two doses of recombinant F protein with interval of 15 days were injected subcutaneously for production of antibodies against the F protein. Antibodies were obtained from serum taken after scarifying the rabbits. Production of polyclonal antibodies was confirmed through Dot Blot Assay and ELISA. In this study maximum antibody titer was estimated to be 4.676 (OD at 450 nm) in case of using concentrated antibody. Produced polyclonal antibodies can be further purified in order to use as tools in biomedical and biochemical researches and diagnostic kits