Purification, characterization and application of recombinant hemagglutinin derived from measles virus

BELIZAN, A.L.; ARGÜELLES, M.H.; BUSOWSKY, I.V.; MANDILE, M.G.; LOZANO, M.E.; TABOGA, O.; GLIKMANN, G.

Mar del Plata, Buenos Aires

Congreso; XLIII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2007

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Introduction: Measles virus, a member of the Paramyxoviridae, is capable of causing acute and persistent infections. The acute infection is followed by life-long immunity in which neutralizing antibodies against the viral haemagglutinin play an important role. The HA protein is a type II surface glycoprotein, and is involved in the attachment to the host cell.   Objectives:Purification and evaluation of recombinant HA protein expressed in baculovirus system and production of polyclonal antibodies. Evaluation of neutralizing antibodies in infected and vaccinated individuals using the recombinant  protein. Methods: A truncated version of Measles HA protein derived from Edmonston strain was expressed using baculovirus system. The expressed HA was purified with Ni-Nta agarose and evaluated by EIA, SDS-PAGE and Western blot. The recombinant HA was used in a capture EIA assay to evaluate neutralizing antibodies. Conclusions:The purified recombinant HA protein was characterized by EIA, SDS-PAGE and Western blot analysis. Results with these methods demonstrated that the protein has been successfully purified from culture supernatants and, as judged by EIA with a conformation similar to the native HA of the MV. Furthermore, application of the recombinant HA in a capture EIA demonstrated the presence of neutralizing IgG antibodies in sera of naturally infected and vaccinated individuals.