Preparation by alkaline treatment and detailed characterisation of empty hepatitis B virus core particles for vaccine and gene therapy applications

Hepatitis B virus (HBV) core (HBc) virus-like particles (VLPs) are one of the most powerful protein engineering tools utilised to expose immunological epitopes and/or cell-targeting signals and for the packaging of genetic material and immune stimulatory sequences. Although HBc VLPs and their numero...

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Vydáno v:Scientific reports Ročník 5; číslo 1; s. 11639
Hlavní autoři: Strods, Arnis, Ose, Velta, Bogans, Janis, Cielens, Indulis, Kalnins, Gints, Radovica, Ilze, Kazaks, Andris, Pumpens, Paul, Renhofa, Regina
Médium: Journal Article
Jazyk:angličtina
Vydáno: London Nature Publishing Group UK 26.06.2015
Nature Publishing Group
Témata:
101/28
631/337/475
639/638/92/612/1256
639/925/352/152
82/29
82/83
Alkalies - chemistry
Amino Acid Sequence
Core particles
Deoxyribonucleic acid
DNA
Electrophoresis, Polyacrylamide Gel
Epitopes
Gene therapy
Genetic Therapy - methods
Glutamic acid
Hepatitis
Hepatitis B
Hepatitis B Core Antigens - genetics
Hepatitis B Core Antigens - immunology
Hepatitis B Core Antigens - metabolism
Hepatitis B virus - genetics
Hepatitis B virus - immunology
Hepatitis B virus - metabolism
Humanities and Social Sciences
Hydrogen-Ion Concentration
Hydrolysis
Lysine
Microscopy, Electron
Molecular Sequence Data
multidisciplinary
Mutation
Nucleic acids
Nucleotide sequence
Packaging
Protein engineering
Ribonucleic acid
RNA
Science
Sequence Homology, Amino Acid
Spectrophotometry, Ultraviolet
Vaccines
Vaccines, Virus-Like Particle - immunology
Virion - genetics
Virion - immunology
Virion - metabolism
Virus Assembly - genetics
Virus Assembly - immunology
Virus-like particles
Yeast
ISSN:2045-2322, 2045-2322
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Shrnutí:Hepatitis B virus (HBV) core (HBc) virus-like particles (VLPs) are one of the most powerful protein engineering tools utilised to expose immunological epitopes and/or cell-targeting signals and for the packaging of genetic material and immune stimulatory sequences. Although HBc VLPs and their numerous derivatives are produced in highly efficient bacterial and yeast expression systems, the existing purification and packaging protocols are not sufficiently optimised and standardised. Here, a simple alkaline treatment method was employed for the complete removal of internal RNA from bacteria- and yeast-produced HBc VLPs and for the conversion of these VLPs into empty particles, without any damage to the VLP structure. The empty HBc VLPs were able to effectively package the added DNA and RNA sequences. Furthermore, the alkaline hydrolysis technology appeared efficient for the purification and packaging of four different HBc variants carrying lysine residues on the HBc VLP spikes. Utilising the introduced lysine residues and the intrinsic aspartic and glutamic acid residues exposed on the tips of the HBc spikes for chemical coupling of the chosen peptide and/or nucleic acid sequences ensured a standard and easy protocol for the further development of versatile HBc VLP-based vaccine and gene therapy applications.
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ISSN:2045-2322
2045-2322
DOI:10.1038/srep11639