title: Purified and highly aggregated chimeric protein DIIIC-2 induces a functional immune response in mice against dengue 2 virus.
abstract: It was previously reported that DIIIC-2 (a fusion protein composed of domain III of the envelope protein and the capsid protein from dengue 2 virus), as an aggregate antigen from a partially purified preparation, induced a functional protective immune response against dengue 2 virus in the mouse encephalitis model. In the present work, a purification procedure was developed for DIIIC-2, and soluble and aggregated fractions of the purified protein were characterized and evaluated in mice. The purification process rendered a protein preparation of 91 % purity, and the remaining 9 % consisted of fragments and aggregates of the same recombinant protein. After the in vitro aggregation process, upon addition of oligodeoxynucleotides, 80 % of the protein formed aggregates, whereas 20 % remained as soluble protein. An immunological evaluation revealed the proper immunogenicity of the aggregated purified protein in terms of induction of antiviral and neutralizing antibodies, cell-mediated immunity and protection upon dengue 2 virus challenge in the mouse encephalitis model. Based on these results, we can assert that the purified protein DIIIC-2 is functional and could be used for further scalable steps and preclinical studies in non-human primates.
authors.affiliation: Vaccine Division, Center for Genetic Engineering and Biotechnology (CIGB), Avenue 31, P.O. Box 6162, Havana 6, 10 600, Cuba.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Marcos
authors.forename: Lázaro
authors.initials: L
authors.lastname: Gil
authors.forename: Laura
authors.initials: L
authors.lastname: Lazo
authors.forename: Alienys
authors.initials: A
authors.lastname: Izquierdo
authors.forename: Enma
authors.initials: E
authors.lastname: Brown
authors.forename: Edith
authors.initials: E
authors.lastname: Suzarte
authors.forename: Iris
authors.initials: I
authors.lastname: Valdés
authors.forename: Angélica
authors.initials: A
authors.lastname: García
authors.forename: Lissandra
authors.initials: L
authors.lastname: Méndez
authors.forename: María G
authors.initials: MG
authors.lastname: Guzmán
authors.forename: Gerardo
authors.initials: G
authors.lastname: Guillén
authors.forename: Lisset
authors.initials: L
authors.lastname: Hermida
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D003715:Dengue / Q000276:immunology* / Q000821:virology; D053059:Dengue Vaccines / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology*; D005260:Female; D006801:Humans; D007111:Immunity, Cellular; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D053059:Dengue Vaccines; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins