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Engineered Recombinant NDV-Fusion Protein and Its Polyclonal Antibodies Production
 

Maira Zahid1, Asma Irshad2, Sana Shakoor1, Tahir Rehman Samiullah1, Naila Shahid1, Adnan Iqbal1, Sana Tanveer1, Shehla J Akram3, M Azam Ali1, Shafique Ahmed1,4, Tayyab Husnain1, Ahmad Ali Shahid1 and Abdul Qayyum Rao1

 
1Centre of Excellence in Molecular Biology, University of the Punjab, 87-West Canal Bank Road Lahore, Pakistan
2University of Management and Technology Lahore Pakistan; 3Akram Medical Complex Lahore, Pakistan
4Allied Health Sciences, The Superior College, Lahore, Pakistan
*Corresponding author: qayyumabdul77@yahoo.com

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.

To Cite This Article: Zahid M, Irshad A, Shakoor S, Samiullah TR, Shahid N, Iqbal A, Tanveer S, Akram SJ, Ali MA, Ahmed S, Husnain T, Shahid AA and Rao AQ, 2020. Engineered recombinant NDV-Fusion protein and its polyclonal antibodies production. Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2020.034

 
   

ISSN 0253-8318 (Print)
ISSN 2074-7764 (Online)



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