Production and purification of recombinant Measles virus hemagglutinin to measure humoral immunity

BELIZAN, A.L.; ARGÜELLES, M.H.; LOZANO, M.E.; TABOGA, O.; GLIKMANN, G.

Puerto Madryn

Congreso; XLVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Measles virus, a member of the Paramyxoviridae, is capable of causing acute and persistent infection. The acute infection is following by life-long immunity in which neutralizing antibodies against the viral haemagglutinin (HA) play an important role. The HA protein is a type II surface glycoprotein, and is involved in the attachment to the host cell. 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 HA protein. A truncated version Measles HA protein (without transmembrane hydrophobic region) derived from Edmonston strain, was expressed using baculovirus System. The HA expressed was purificated with Ni-Nta agarose .The purified HA was evaluated by EIA, SDS-PAGE and western blot. The recombinant HA was used in capture EIA assay to evaluate neutralizing antibodies. The EIA results were compared to those obtained with a micro-neutralization assay. The purified recombinant HA protein was detected by EIA SDSPAGE and Western blot. Moreover, recombinant HA was detected by EIA using human serum from infected and vaccinated individual. These results suggest that the recombinant HA can be used in evaluation of immunological parameters