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Production of Chimeric Hepatitis B Virus Surface Antigens in Mammalian Cells.
Dobrica, Mihaela-Olivia; Lazar, Catalin; Branza-Nichita, Norica.
Affiliation
  • Dobrica MO; Institute of Biochemistry of the Romanian Academy, Bucharest, Romania.
  • Lazar C; Institute of Biochemistry of the Romanian Academy, Bucharest, Romania.
  • Branza-Nichita N; Institute of Biochemistry of the Romanian Academy, Bucharest, Romania. nichita@biochim.ro.
Methods Mol Biol ; 2183: 83-94, 2021.
Article de En | MEDLINE | ID: mdl-32959242
ABSTRACT
The small (S) envelope protein of the Hepatitis B Virus (HBV), HBV-S, has the unique ability to self-assemble into highly immunogenic subviral particles (SVPs), in the absence of other viral factors, in eukaryotic cells, including those of nonhepatic origin. This feature is currently exploited for generation of SVPs exposing heterologous epitopes on their surface that can be used as vaccine candidates to target various diseases. Here, we describe a simple and robust method for production of such chimeric HBV-S protein-based SVPs in transiently transfected HEK293T cells and purification from cell supernatants by ultracentrifugation on sucrose cushion and sucrose step gradients. The SVPs obtained by this methodology have been successfully used in immunogenicity studies in animal models.
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Texte intégral: 1 Collection: 01-internacional Base de données: MEDLINE Sujet principal: Protéines de fusion recombinantes / Techniques de culture cellulaire en batch / Antigènes de surface du virus de l'hépatite B Type d'étude: Prognostic_studies Limites: Animals / Humans Langue: En Journal: Methods Mol Biol Sujet du journal: BIOLOGIA MOLECULAR Année: 2021 Type de document: Article Pays d'affiliation: Roumanie

Texte intégral: 1 Collection: 01-internacional Base de données: MEDLINE Sujet principal: Protéines de fusion recombinantes / Techniques de culture cellulaire en batch / Antigènes de surface du virus de l'hépatite B Type d'étude: Prognostic_studies Limites: Animals / Humans Langue: En Journal: Methods Mol Biol Sujet du journal: BIOLOGIA MOLECULAR Année: 2021 Type de document: Article Pays d'affiliation: Roumanie