دورية أكاديمية
Recombinant Expression of Tandem-HBc Virus-Like Particles (VLPs).
| العنوان: | Recombinant Expression of Tandem-HBc Virus-Like Particles (VLPs). |
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| المؤلفون: | Stephen SL; School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK.; CPI, National Biologics Manufacturing Centre, Darlington, UK., Beales L; Mologic, Bedfordshire, UK., Peyret H; Department of Biological Chemistry, John Innes Centre, Norwich, UK., Roe A; School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK., Stonehouse NJ; School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK., Rowlands DJ; School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK. d.j.rowlands@leeds.ac.uk. |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1776, pp. 97-123. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | Hepatitis B Core Antigens/*genetics , Hepatitis B virus/*genetics , Recombinant Fusion Proteins/*genetics , Vaccines, Virus-Like Particle/*genetics, Amino Acid Sequence ; Bacteria/virology ; Epitopes/genetics ; Pichia/genetics ; Pichia/virology ; Plants/virology ; Viral Vaccines/genetics ; Yeasts/virology |
| مستخلص: | The hepatitis B virus (HBV) core protein (HBc) has formed the building block for virus-like particle (VLP) production for more than 30 years. The ease of production of the protein, the robust ability of the core monomers to dimerize and assemble into intact core particles, and the strong immune responses they elicit when presenting antigenic epitopes all demonstrate its promise for vaccine development (reviewed in Pumpens and Grens (Intervirology 44: 98-114, 2001)). HBc has been modified in a number of ways in attempts to expand its potential as a novel vaccine platform. The HBc protein is predominantly α-helical in structure and folds to form an L-shaped molecule. The structural subunit of the HBc particle is a dimer of monomeric HBc proteins which together form an inverted T-shaped structure. In the assembled HBc particle the four-helix bundle formed at each dimer interface appears at the surface as a prominent "spike." The tips of the "spikes" are the preferred sites for the insertion of foreign sequences for vaccine purposes as they are the most highly exposed regions of the assembled particles. In the tandem-core modification two copies of the HBc protein are covalently linked by a flexible amino acid sequence which allows the fused dimer to fold correctly and assemble into HBc particles. The advantage of the modified structure is that the assembly of the dimeric subunits is defined and not formed by random association. This facilitates the introduction of single, larger sequences at the tip of each surface "spike," thus overcoming the conformational clashes contingent on insertion of large structures into monomeric HBc proteins.Differences in inserted sequences influence the assembly characteristics of the modified proteins, and it is important to optimize the design of each novel construct to maximize efficiency of assembly into regular VLPs. In addition to optimization of the construct, the expression system used can also influence the ability of recombinant structures to assemble into regular isometric particles. Here, we describe the production of recombinant tandem-core particles in bacterial, yeast and plant expression systems. |
| فهرسة مساهمة: | Keywords: Escherichia coli expression; HBc VLPs; Hepatitis B core (HBc); Pichia pastoris expression; Plant expression |
| المشرفين على المادة: | 0 (Epitopes) 0 (Hepatitis B Core Antigens) 0 (Recombinant Fusion Proteins) 0 (Vaccines, Virus-Like Particle) 0 (Viral Vaccines) |
| تواريخ الأحداث: | Date Created: 20180606 Date Completed: 20190204 Latest Revision: 20190215 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1007/978-1-4939-7808-3_7 |
| PMID: | 29869237 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-6029 |
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| DOI: | 10.1007/978-1-4939-7808-3_7 |