CAS OpenIR
In vitro preparation of uniform and nucleic acid free hepatitis B core particles through an optimized disassembly-purification-reassembly process
Zhang, Yao1,2; Liu, Yongdong1; Zhang, Bingyang1,3; Yin, Shuang1,3; Li, Xiunan1; Zhao, Dawei1; Wang, Weiying1; Bi, Jingxiu3; Su, Zhiguo1
2021-02-01
Source PublicationPROTEIN EXPRESSION AND PURIFICATION
ISSN1046-5928
Volume178Pages:7
AbstractStructure 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.
KeywordHepatitis B core (HBc) Uniform VLPs Nucleic acid removal Disassembly-purification-reassembly
DOI10.1016/j.pep.2020.105747
Language英语
WOS KeywordVIRUS-LIKE PARTICLES ; STABILITY ; DELIVERY ; VACCINES ; EXPRESSION
Funding ProjectNational Natural Science Foundation of China[21576267]
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
Funding OrganizationNational Natural Science Foundation of China
WOS IDWOS:000651432900002
PublisherACADEMIC PRESS INC ELSEVIER SCIENCE
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/48818
Collection中国科学院过程工程研究所
Corresponding AuthorLiu, Yongdong; Su, Zhiguo
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Univ Adelaide, Sch Chem Engn & Adv Mat, Adelaide, SA 5005, Australia
Recommended Citation
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.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
Google Scholar
Similar articles in Google Scholar
[Zhang, Yao]'s Articles
[Liu, Yongdong]'s Articles
[Zhang, Bingyang]'s Articles
Baidu academic
Similar articles in Baidu academic
[Zhang, Yao]'s Articles
[Liu, Yongdong]'s Articles
[Zhang, Bingyang]'s Articles
Bing Scholar
Similar articles in Bing Scholar
[Zhang, Yao]'s Articles
[Liu, Yongdong]'s Articles
[Zhang, Bingyang]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.