Session:

Vaccines & Vaccine Development

Abstract No.:

48.018

Title:

Baculovirus system as a protein expression of influenza A/H1N1p virus

Author(s):

E. Loza-Rubio¹, E. Rojas-Anaya¹, C. Chevalier², E. Rangel-Huerta¹, B. Da Costa², B. Delmas²; ¹INIFAP, National Center of Research in Animal Microbiology , Mexico, MEXICO CITY/MX, ²Institut National de la Recherche Agronomique, Unité de Virologie et Immunologie Moléculaires, Jouy-en-Josas/FR

Abstract:

Background: Influenza is a zoonotic disease that has caused major epidemics in the world in both humans and animals causing economic losses. Influenza A/H1N1/2009 virus is circulating among human and animal populations. It is, therefore, important to develop strategies to produce protective antibody responses against H1N1 virus. There have been international efforts to obtain and a universal vaccine for different subtypes of the virus. Proteins that can be used to achieve this purpose are structural. So the aim of this study was to obtain the M and NS1 recombinant proteins in addition to the HA1 of influenza virus strain A/H1N1/2009 (Paris 2009) using baculovirus systems for used as vaccine and diagnostic purposes
Methods: The genes M1, NS1, HA and NP influenza virus A/H1N1/Paris/2009 were amplified by RT-PCR and were subcloned separately into the vector pST-28 and subsequently cloned into the pFastBac vector to obtain four recombinant baculovirus with which separately Sf9 insect cells were infected. Insect cells extracts were used to identify proteins by western blot using monoclonal antibodies and a pool of sera from people infected with the virus.
Results: In the western blot were recognized a protein of ~26 kDa (M protein), a ~26 kDa (NS1 protein) and another of ~65 kDa (HA1 protein). This monoclonal antibody using both patient sera thereby demonstrated that these proteins retained their antigenic properties.
Conclusion: We conclude that the baculovirus expression system was efficient for obtaining these four recombinant proteins of influenza virus. These products could be used as immunogen and/or reactive for ELISA test.