title: Expression of a secreted version of the hemagglutinin-neuraminidase glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
abstract: The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) constitutes, together with the fusion glycoprotein, the main surface antigen of this avian pathogen, which causes a highly contagious disease, relevant economically worldwide. The purpose of this work was to obtain the HN glycoprotein as a soluble antigen in culture supernatants of recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells and to evaluate its application to the development of a recombinant enzyme-linked immunosorbent assay (rELISA) for the analysis of chicken sera. A transfer vector for baculovirus containing the sequence of a melittin signal peptide was constructed and the sequence coding for HN protein without its own signal peptide was cloned. The recombinant protein was secreted and recovered easily from the culture medium of Sf9-infected cells. The recombinant protein was evaluated as antigen for ELISA coating the plates with the recovered HN using 79 positive and 142 negative samples. The Cohen kappa value resulted 0.91, indicating excellent agreement between the rELISA and the hemagglutinin inhibition tests. The rELISA was also compared with a commercial ELISA, finding high levels of agreement between both assays. The present results show that the cloning strategy developed yielded the HN protein free in the cell culture supernatant and that the recombinant protein retained its reactivity with anti-NDV HN antibodies in chicken sera.
authors: Instituto de Biotecnología, Centro de Investigación en Ciencias Veterinarias y Agronómicas, Instituto Nacional de Tecnologia Agropecuaria, Castelar, Buenos Aires, Argentina.
authors: Silvina Chimeno
authors: SC
authors: Zoth
authors: Evangelina
authors: E
authors: Gómez
authors: Juan Manuel
authors: JM
authors: Carballeda
authors: Elisa
authors: E
authors: Carrillo
authors: Analía
authors: A
authors: Berinstein
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured / Q000821:virology; D002645:Chickens / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005822:Genetic Vectors / Q000235:genetics; D015707:HN Protein / Q000096:biosynthesis*; D006385:Hemagglutination Inhibition Tests / Q000662:veterinary; D009521:Newcastle Disease / Q000175:diagnosis*; D009522:Newcastle disease virus / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D018411:Spodoptera / Q000821:virology
chemical_list: D015707:HN Protein; D011994:Recombinant Proteins