title: Generation and characterization of potential dengue vaccine candidates based on domain III of the envelope protein and the capsid protein of the four serotypes of dengue virus.
abstract: Dengue is currently one of the most important arthropod-borne diseases, causing up to 25,000 deaths annually. There is currently no vaccine to prevent dengue virus infection, which needs a tetravalent vaccine approach. In this work, we describe the cloning and expression in Escherichia coli of envelope domain III-capsid chimeric proteins (DIIIC) of the four dengue serotypes as a tetravalent dengue vaccine candidate that is potentially able to generate humoral and cellular immunity. The recombinant proteins were purified to more than 85 % purity and were recognized by anti-dengue mouse and human sera. Mass spectrometry analysis verified the identity of the proteins and the correct formation of the intracatenary disulfide bond in the domain III region. The chimeric DIIIC proteins were also serotype-specific, and in the presence of oligonucleotides, they formed aggregates that were visible by electron microscopy. These results support the future use of DIIIC recombinant chimeric proteins in preclinical studies in mice for assessing their immunogenicity and efficacy.
authors.affiliation: Center for Genetic Engineering and Biotechnology (CIGB), Avenue 31, P.O. Box 6162, Havana 6, 10600, Cuba.
authors.forename: Edith
authors.initials: E
authors.lastname: Suzarte
authors.forename: Ernesto
authors.initials: E
authors.lastname: Marcos
authors.forename: Lázaro
authors.initials: L
authors.lastname: Gil
authors.forename: Iris
authors.initials: I
authors.lastname: Valdés
authors.forename: Laura
authors.initials: L
authors.lastname: Lazo
authors.forename: Yassel
authors.initials: Y
authors.lastname: Ramos
authors.forename: Yusleidi
authors.initials: Y
authors.lastname: Pérez
authors.forename: Viviana
authors.initials: V
authors.lastname: Falcón
authors.forename: Yaremis
authors.initials: Y
authors.lastname: Romero
authors.forename: María G
authors.initials: MG
authors.lastname: Guzmán
authors.forename: Sirenia
authors.initials: S
authors.lastname: González
authors.forename: Juan
authors.initials: J
authors.lastname: Kourí
authors.forename: Gerardo
authors.initials: G
authors.lastname: Guillén
authors.forename: Lisset
authors.initials: L
authors.lastname: Hermida
mesh_terms: D000956:Antigens, Viral / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D053059:Dengue Vaccines; D003716:Dengue Virus / Q000145:classification* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004926:Escherichia coli; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000276:immunology; D012703:Serotyping; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D053059:Dengue Vaccines; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins