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A Competent Hepatocyte Model Examining Hepatitis B Virus Entry through Sodium Taurocholate Cotransporting Polypeptide as a Therapeutic Target
Published on: May 10, 2022
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Cost-effective purification process development for chimeric hepatitis B core (HBc) virus-like particles assisted by
Bingyang Zhang1, Shuang Yin1, Yingli Wang2
1School of Chemical Engineering & Advanced Materials, Faculty of Engineering, Computer and Mathematical Sciences University of Adelaide Adelaide SA Australia.
Engineering in Life Sciences
|June 18, 2021
Summary
A cost-effective purification process was developed for chimeric hepatitis B core (HBc) virus-like particles (VLPs) displaying foreign epitopes. Molecular dynamic simulations guided optimization, ensuring VLP structural integrity and bioactivity post-purification.
Area of Science:
- Biotechnology
- Structural Biology
- Virology
Background:
- Engineering virus-like particles (VLPs) with foreign epitopes presents purification challenges due to altered molecular conformation.
- Hepatitis B core (HBc) VLPs are a versatile platform for displaying heterologous antigens.
Purpose of the Study:
- To develop a cost-effective purification strategy for chimeric HBc VLPs displaying Epstein-Barr nuclear antigen 1 (EBNA1) and hepatitis C virus (HCV) core antigens.
- To assess the impact of foreign epitope insertion on VLP stability and structure.
Main Methods:
- Expression of chimeric VLPs in *Escherichia coli*.
- Molecular dynamic (MD) simulations to predict VLP stability and hydrophobicity.
- Optimization of ammonium sulfate precipitation for host cell protein removal.
- POROS 50 HQ chromatography for DNA impurity removal.
- Characterization using tertiary structure analysis, transmission electron microscopy (TEM), and immunogenicity assays.
Main Results:
- Chimeric VLPs were expressed in soluble form with high yields.
- MD simulations indicated higher hydrophobicity and lower stability for HCV core-HBc compared to EBNA1-HBc.
- Optimized ammonium sulfate precipitation and chromatography achieved 99% DNA impurity removal.
- HCV core epitope insertion reduced the α-helix ratio, contributing to decreased stability.
- Purified chimeric VLPs maintained VLP structure and bioactivity.
Conclusions:
- A cost-effective purification process was successfully developed for chimeric HBc VLPs.
- MD simulations are valuable tools for predicting VLP behavior and guiding purification strategies.
- The developed method ensures the structural integrity and bioactivity of engineered VLPs for potential therapeutic applications.
