Baculovirus expression and purification of peste-des-petits-ruminants virus nucleocapsid protein and its application in diagnostic assay

Publication date: Available online 17 July 2018Source: BiologicalsAuthor(s): Suresh H. Basagoudanavar, Madhusudan Hosamani, D. Muthuchelvan, R.P. Singh, R. Santhamani, B.P. Sreenivasa, P. Saravanan, A.B. Pandey, R.K. Singh, Ramamurthy VenkataramananAbstractPeste-des-petits-ruminants (PPR) is a contagious and highly devastating disease of small ruminants. For control of endemic PPR, adequate supply of affordable and reliable diagnostics is critical for effective surveillance, along with the use of highly efficacious live vaccines that are currently available. The nucleocapsid (N) protein of PPR virus (PPRV) is an important candidate antigen for developing specific diagnostic, as it is a major viral protein being highly immunogenic and conserved among the structural proteins. In the present study, we expressed the N protein of PPRV (Sungri/96 strain), in baculovirus expression system and purified using affinity column chromatography. The recombinant protein reacted well with PPRV anti-N monoclonal antibodies and PPRV-specific polyclonal antiserum, suggesting that the expressed protein was authentic and in native form. The recombinant protein was evaluated as antigen in the diagnostic ELISA as reference positive control in place of whole virus antigen. The utility of recombinant PPRV N protein circumvents the need to use live PPRV antigen in the routinely used diagnostics targeting ‘N’ protein of PPRV, thus allowing large-scale field application of the test.

Source: Biologicals - July 18, 2018 Category: Biology Source Type: research