IIBBA   05544
INSTITUTO DE INVESTIGACIONES BIOQUIMICAS DE BUENOS AIRES
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Cloning, expression and purification of HCV Core variant fused to the carrier protein BLS
Autor/es:
MARIACARLA DAUSA DE ARMAS; GILLIAN MARTINEZ DONATO; ROMINA PARDO; GISELLE GHERSI; FERNANDO A. GOLDBAUM; SANTIAGO DUEÑAS CARRERA
Lugar:
Varadero
Reunión:
Congreso; Congreso Hepatitis 2014; 2014
Resumen:
The Lumazine syntethase enzyme is involved in the synthesis of riboflavin in Brucella spp (BLS). BLS is a highly immunogenic stable decamer, successfully used as a carrier for several proteins. BLS immunization activates murine dendritic cells (DC) and recruits DC, B cells, CD8+ and CD4+ cells at the draining lymph nodes (LN) via TLR4. BLS induces the cross-presentation of associated peptides and a TLR4-dependent specific cytotoxicity, being potentially useful for the development of subunits vaccines against viral infections. In our project, a fragment of hepatitis C virus Core structural protein (amino acids 1- 145) was cloned in pET11c plasmid fused to the amino terminal end of BLS. The fusion Core-BLS protein was obtained in E. coli strain BL21 RIPL as inclusion body. Core-BLS was identified as a 35 kDa polypeptide by immunoblot analysis using anti-BLS monoclonal antibody (mAb) and anti-Core mAb. Co-BLS was purified by washed pellet, and cationic exchange chromatography using SP Sepharose. These results show that Co-BLS chimeric protein is succefully produced by E. coli and could be used to carry out immunogenicity experiments to evaluate specific immune response against HCV Core protein in animal models.