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J Nanobiotechnology. 2018 Apr 13;16(1):39. doi: 10.1186/s12951-018-0363-0.

Enhanced stability of a chimeric hepatitis B core antigen virus-like-particle (HBcAg-VLP) by a C-terminal linker-hexahistidine-peptide.

Author information

1
Biopharmaceutical New Technologies (BioNTech) Protein Therapeutics Corporation, An der Goldgrube 12, 55131, Mainz, Germany.
2
Department of Internal Medicine III, Translational and Experimental Oncology, University Medical Center of Johannes Gutenberg University, Langenbeckstrasse 1, 55131, Mainz, Germany.
3
TRON Translational Oncology, University Medical Center of Johannes Gutenberg University, TRON gGmbH, Freiligrathstrasse 12, 55131, Mainz, Germany.
4
Institute of Zoology, Johannes Gutenberg University, Johannes-von-Müller-Weg 6, 55128, Mainz, Germany.
5
Anatomical Institute, Otto-Hahn Platz 8, 24118, Kiel, Germany.
6
Ganymed Pharmaceuticals AG, An der Goldgrube 12, 55131, Mainz, Germany.
7
Biopharmaceutical New Technologies (BioNTech) Protein Therapeutics Corporation, An der Goldgrube 12, 55131, Mainz, Germany. sahin@uni-mainz.de.
8
Department of Internal Medicine III, Translational and Experimental Oncology, University Medical Center of Johannes Gutenberg University, Langenbeckstrasse 1, 55131, Mainz, Germany. sahin@uni-mainz.de.
9
TRON Translational Oncology, University Medical Center of Johannes Gutenberg University, TRON gGmbH, Freiligrathstrasse 12, 55131, Mainz, Germany. sahin@uni-mainz.de.

Abstract

BACKGROUND:

Virus-like-particles (VLPs) are attractive nanoparticulate scaffolds for broad applications in material/biological sciences and medicine. Prior their functionalization, specific adaptations have to be carried out. These adjustments frequently lead to disordered particles, but the particle integrity is an essential factor for the VLP suitability. Therefore, major requirements for particle stabilization exist. The objective of this study was to evaluate novel stabilizing elements for functionalized chimeric hepatitis B virus core antigen virus-like particles (HBcAg-VLP), with beneficial characteristics for vaccine development, imaging or delivery.

RESULTS:

The effects of a carboxy-terminal polyhistidine-peptide and an intradimer disulfide-bridge on the stability of preclinically approved chimeric HBcAg-VLPs were assessed. We purified recombinant chimeric HBcAg-VLPs bearing different modified C-termini and compared their physical and chemical particle stability by quantitative protein-biochemical and biophysical techniques. We observed lower chemical resistance of T = 3- compared to T = 4-VLP (triangulation number) capsids and profound impairment of accessibility of hexahistidine-peptides in assembled VLPs. Histidines attached to the C-terminus were associated with superior mechanical and/or chemical particle stability depending on the number of histidine moieties. A molecular modeling approach based on cryo-electron microscopy and biolayer interferometry revealed the underlying structural mechanism for the strengthening of the integrity of VLPs. Interactions triggering capsid stabilization occur on a highly conserved residue on the basis of HBcAg-monomers as well as on hexahistidine-peptides of adjacent monomers. This new stabilization mechanism appears to mimic an evolutionary conserved stabilization concept for hepadnavirus core proteins.

CONCLUSIONS:

These findings establish the genetically simply transferable C-terminal polyhistidine-peptide as a general stabilizing element for chimeric HBcAg-VLPs to increase their suitability.

KEYWORDS:

Bioengineering; Bionanoparticles; Capsid and particle stability; Chimeric VLP; Computational biology; Hepatitis B virus; Molecular mechanism; Structural biology; Vaccine; Virus-like-particles (VLP)

PMID:
29653575
PMCID:
PMC5897928
DOI:
10.1186/s12951-018-0363-0
[Indexed for MEDLINE]
Free PMC Article

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