In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process

doi:10.1016/j.pep.2020.105747

	In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process
	Zhang, Yao 1,3; Liu, Yongdong 1; Zhang, Bingyang 1,2; Yin, Shuang 1,2; Li, Xiunan 1; Zhao, Dawei 1; Wang, Weiying 1; Bi, Jingxiu 2; Su, Zhiguo 1
刊名	PROTEIN EXPRESSION AND PURIFICATION
	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.
资助项目	National Natural Science Foundation of China[21576267]
WOS关键词	VIRUS-LIKE PARTICLES ; STABILITY ; DELIVERY ; VACCINES ; EXPRESSION
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000651432900002
资助机构	National Natural Science Foundation of China
内容类型	期刊论文
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.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.Univ Adelaide, Sch Chem Engn & Adv Mat, Adelaide, SA 5005, Australia 3.Univ Chinese Acad Sci, Beijing 100049, 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.