Cost‐effective purification process development for chimeric hepatitis B core (HBc) virus‐like particles assisted by molecular dynamic simulation

• Published in: Engineering in life sciences Vol. 21; no. 6; pp. 438 - 452
• Main Authors: Zhang, Bingyang, Yin, Shuang, Wang, Yingli, Su, Zhiguo, Bi, Jingxiu
• Format: Journal Article
• Language: English
• Published: Weinheim John Wiley & Sons, Inc 01.06.2021; John Wiley and Sons Inc; Wiley-VCH
• Subjects: Ammonium; Ammonium sulfate; Ammonium sulphate; Antigenic determinants; Antigens; Bacteria; Biological activity; Cancer; Chromatography; Comparative analysis; Dynamic stability; E coli; Economic aspects; Epitopes; Epstein-Barr virus; Escherichia coli; Hepatitis B; hepatitis B core; Hepatitis C; Hepatitis C virus; Hydrophobicity; Immunogenicity; Impurities; Infections; Molecular conformation; molecular dynamic simulation; Molecular dynamics; Molecular weight; protein characterization; protein purification; Protein structure; Proteins; Purification; Simulation; Tertiary structure; Transmission electron microscopy; Vaccines; Viral infections; Viruses; virus‐like particle; United States > US; China
• ISSN: 1618-0240; 1618-2863
• DOI: 10.1002/elsc.202000104

Inserting foreign epitopes to hepatitis B core (HBc) virus‐like particles (VLPs) could influence the molecular conformation and therefore vary the purification process. In this study, a cost‐effective purification process was developed for two chimeric HBc VLPs displaying Epstein–Barr nuclear antigens 1 (EBNA1), and hepatitis C virus (HCV) core. Both chimeric VLPs were expressed in soluble form with high production yields in Escherichia coli. Molecular dynamic (MD) simulation was employed to predict the stability of chimeric VLPs. HCV core‐HBc was found to be less stable in water environment compared with EBNA1‐HBc, indicating its higher hydrophobicity. Assisting with MD simulation, ammonium sulfate precipitation was optimized to remove host cell proteins with high target protein recovery yields. Moreover, 99% DNA impurities were removed using POROS 50 HQ chromatography. In characterization measurement, we found that inserting HCV core epitope would reduce the ratio of α‐helix of HCV core‐HBc. This could be another reason on the top of its higher hydrophobicity predicted by MD simulation, causing its less stability. Tertiary structure, transmission electron microscopy, and immunogenicity results indicate that two chimeric VLPs maintained correct VLP structure ensuring its bioactivity after being processed by the developed cost‐effective purification approach.