Single Step Detergent Assisted Extraction and Solubilization of the Recombinant Matrix Protein of Newcastle Disease Virus (NDV) and Development of ELISA for Detection of Anti-NDV Antibodies in Chickens
Single Step Detergent Assisted Extraction and Solubilization of the Recombinant Matrix Protein of Newcastle Disease Virus (NDV) and Development of ELISA for Detection of Anti-NDV Antibodies in Chickens
Zahra Naz1,2, Fouzia Ismat1, Muhammad Saleem2, Mazhar Iqbal1, Aamir Shehzad1 and Moazur Rahman1,2*
ABSTRACT
The matrix (M) protein is the most abundant structural protein in Newcastle disease virus (NDV), the causative agent of Newcastle disease (ND) in chickens. Owing to its highly conserved nature among NDV strains and also due to its pivotal role in the viral life cycle, the M protein can be employed as a promising diagnostic antigen for reliable detection of NDV infection in chickens. In the present study, we have devised a strategy for extraction and solubilization of the NDV M protein from Escherichia coli in a single step using a non-ionic detergent, lauryl-dimethylamine oxide (LDAO), enabling the purification of the detergent-solubilized M protein in a soluble form through affinity chromatography without compromising the structural integrity of the protein. Using the purified M protein as a diagnostic antigen, an indirect enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of anti-NDV antibodies in multiple serum samples collected from different poultry farms of district Faisalabad, Pakistan. The data presented here reveal that the recombinant detergent-solubilized M protein is an active, promising diagnostic antigen and can be exploited in an indirect ELISA for rapid and reliable detection of anti-NDV antibodies in chickens. Owing to the conserved nature of the M protein and the ease of extraction and solubilization in a single step using LDAO, the developed ELISA can be up-scaled for rapid and reliable detection of anti-NDV antibodies and/or NDV infection in a large number of serum samples collected from chickens originating from different sources (poultry farms).
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