In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process
文献类型：期刊论文
| 作者 | Zhang, Yao2,3; Liu, Yongdong3; Zhang, Bingyang1,3; Yin, Shuang1,3; Li, Xiunan3; Zhao, Dawei3; Wang, Weiying3; Bi, Jingxiu1; Su, Zhiguo3 |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2021-02-01 |
| 卷号 | 178页码:7 |
| 关键词 | Hepatitis B core (HBc) Uniform VLPs Nucleic acid removal Disassembly-purification-reassembly |
| ISSN号 | 1046-5928 |
| DOI | 10.1016/j.pep.2020.105747 |
| 英文摘要 | Structure heterogeneity and host nucleic acids contamination are two major problems for virus-like particles (VLPs) produced by various host cells. In this study, an in vitro optimized disassembly-purification-reassembly process was developed to obtain uniform and nucleic acid free hepatitis B core (HBc) based VLPs from E. coli fermentation. The process started with ammonium sulfate precipitation of all heterogeneous HBc structures after cell disintegration. Then, dissolution and disassembly of pellets into basic subunits were carried out under the optimized disassembly condition. All contaminants, including host nucleic acids and proteins, were efficiently removed with affinity chromatography. The purified subunits reassembled into VLPs by final removal of the chaotmpic agent. Two uniform and nucleic acid free HBc-based VLPs, truncated HBc(149) and chimeric HBc(183)-MAGE3 I, were successfully prepared. It was found that disassembly degree of HBc-based VLPs had a great influence on the protein yield, nucleic acid removal and reassembly efficiency. 4 M urea was optimal because lower concentration would not disassemble the particles completely while higher concentration would further denature the subunits into disordered aggregate and could not be purified and reassembled efficiently. For removal of strong binding nucleic acids such as in the case of HBc(183)-MAGE3 I, benzonase nuclease was added to the disassembly buffer before affinity purification. Through the optimized downstream process, uniform and nucleic acid free HBc(149) VLPs and HBc(183)-MAGE3 I VLPs were obtained with purities above 90% and yields of 55.2 and 43.0 mg/L, respectively. This study would be a reference for efficient preparation of other VLPs. |
| WOS关键词 | VIRUS-LIKE PARTICLES ; STABILITY ; DELIVERY ; VACCINES ; EXPRESSION |
| 资助项目 | National Natural Science Foundation of China[21576267] |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 语种 | 英语 |
| WOS记录号 | WOS:000651432900002 |
| 出版者 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| 资助机构 | National Natural Science Foundation of China |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/48817]  |
| 专题 | 中国科学院过程工程研究所 |
| 通讯作者 | Liu, Yongdong; Su, Zhiguo |
| 作者单位 | 1.Univ Adelaide, Sch Chem Engn & Adv Mat, Adelaide, SA 5005, Australia 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China |
| 推荐引用方式 GB/T 7714 | Zhang, Yao,Liu, Yongdong,Zhang, Bingyang,et al. In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process[J]. PROTEIN EXPRESSION AND PURIFICATION,2021,178:7. |
| APA | Zhang, Yao.,Liu, Yongdong.,Zhang, Bingyang.,Yin, Shuang.,Li, Xiunan.,...&Su, Zhiguo.(2021).In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process.PROTEIN EXPRESSION AND PURIFICATION,178,7. |
| MLA | Zhang, Yao,et al."In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process".PROTEIN EXPRESSION AND PURIFICATION 178(2021):7. |
入库方式： OAI收割
来源：过程工程研究所
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