title: Production of Monoclonal Antibody Against Recombinant Polypeptide From the Erns Coding Region of the Bovine Viral Diarrhea Virus.
abstract: BACKGROUND
  Bovine viral diarrhea (BVD) is an economically important cattle disease with a worldwide distribution. Detection and elimination of animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) is essential for the control of BVD and eradication of BVDV. There are usually no pathognomonic clinical signs of BVDV infection. Diagnostic investigations therefore rely on laboratory-based detection of the virus, or virus-induced antigens or antibodies.
  
  
  OBJECTIVE
  Erns as an immunogenic protein of BVDV, is genetically and antigenically conserved among different isolates and therefore, is a candidate antigen for development of the enzyme linked immunosorbent assay (ELISA) for serological studies or identification of PI animals. The aim of this study was to produce a monoclonal antibody (MAb) against recombinant Erns.
  
  
  METHODS
  For this purpose, recombinant maltose-binding protein (MBP)-Erns protein was expressed in Escherichia coli and purified using amylose resin chromatography column and used as an antigen in MAb production. Spleen cells of the immunized mice with the recombinant antigen were fused with SP2/0 myeloma cells. Next, culture supernatants of primary clones of fused cells were screened by indirect ELISA. After three rounds of cloning, the reactivity of the MAbs with recombinant and natural antigen was established by Western blotting.
  
  
  RESULTS
  Based on our results, MAb against recombinant Erns was produced and reacted successfully with recombinant and natural antigens.
  
  
  CONCLUSIONS
  With regards to the role of Erns in the identification of PI animals, it appears that Erns recombinant antigen and the specific monoclonal antibodies produced against it may be suitable for developing BVDV laboratory diagnostic assays.
authors: Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, IR Iran.
authors: Masood Reza
authors: MR
authors: Seyfi Abad Shapouri
authors: Department of Pharmacognosy, Faculty of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran.
authors: Maryam
authors: M
authors: Ekhtelat
authors: Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, IR Iran.
authors: Masood
authors: M
authors: Ghorbanpoor Najaf Abadi
authors: Department of Pathobiology, Faculty of Para Veterinary Science, Bu-Ali Sina University, Hamedan, IR Iran.
authors: Pezhman
authors: P
authors: Mahmoodi Koohi
authors: Department of Quality Control, Razi Vaccine and Serum Research Institute, Karaj, IR Iran.
authors: Mohsen
authors: M
authors: Lotfi
keywords: Bovine Viral Diarrhea; Monoclonal Antibody; Recombinant Erns