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

被引:6
作者
Zhang, Bingyang [1 ]
Yin, Shuang [1 ]
Wang, Yingli [2 ]
Su, Zhiguo [3 ]
Bi, Jingxiu [1 ]
机构
[1] Univ Adelaide, Fac Engn Comp & Math Sci, Sch Chem Engn & Adv Mat, Adelaide, SA 5005, Australia
[2] Shanxi Univ Tradit Chinese Med, Sch Chinese Med & Food Engn, Jinzhong, Shanxi, Peoples R China
[3] Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing, Peoples R China
来源
ENGINEERING IN LIFE SCIENCES | 2021年 / 21卷 / 06期
基金
中国国家自然科学基金;
关键词
hepatitis B core; molecular dynamic simulation; protein characterization; protein purification; virus‐ like particle; INDUCED EPITOPIC SUPPRESSION; ESCHERICHIA-COLI; TRYPTOPHAN FLUORESCENCE; VACCINE; SURFACE; PROTEINS; CARRIER; CHROMATOGRAPHY; GENERATION; EXPRESSION;
D O I
10.1002/elsc.202000104
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 alpha-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.
引用
收藏
页码:438 / 452
页数:15
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