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A two-step heat treatment of cell disruption supernatant enables efficient removal of host cell proteins before chromatographic purification of HBc particles
Li, Zhengjun1,2; Wei, Jiangxue1,2; Yang, Yanli1; Liu, Lili1; Ma, Guanghui1; Zhang, Songping1; Su, Zhiguo1
2018-12-21
Source PublicationJOURNAL OF CHROMATOGRAPHY A
ISSN0021-9673
Volume1581Pages:71-79
Abstract

The thermal stability of HBc particles was systematically investigated for efficient removal of host cell proteins (HCP) by heat treatment before chromatographic step. The HBc particles were found stable up to 80 degrees C for 30 min without any noticeable change in circular dichroism spectra, fluorescence spectra and transmission electron microscope observation. When heating was applied to precipitate the HCP in the cell disruption supernatant of HBc fermentation, the HCP removal effect was more obvious as the temperature went higher. However, a phenomenon was found beyond 70 degrees C where the recovered HBc particles had larger than normal size and molecular weight as observed by dynamic light scattering and multi-angle laser light scattering. Analysis found that the HBc particles possess nanopores which expand with temperature. When the temperature was above 70 degrees, the pores were large enough for some HCP to penetrate in, but not being able to get out after cooling down. To fully utilize the thermal stability and avoid the interference of HCP entering, a two-step heat treatment strategy was designed. The supernatant was firstly heated up to 60 degrees C for 30 min to precipitate most HCP, then another 30 min at 70 degrees C was used to remove the rest impurities. The two-step heat treatment effectively avoided the HCP entering problem, achieving 85.8% particle recovery and 74.7% purity. With further one-step hydrophobic interaction chromatography, the purity was increased to 99.0% with overall process recovery of 77.7%, considerably higher than those reported in the literature. The same process design was applied to purify three HBc-related products, including OVA-HBc, M2e-HBc and NP-HBc. All recoveries were higher than 50% with purity greater than 97%. (C) 2018 Elsevier B.V. All rights reserved.

KeywordHepatitis b Core Protein Thermal Stability Particles Purification Host Cell Protein Chromatography
DOI10.1016/j.chroma.2018.10.050
Language英语
WOS KeywordVirus Core Particles ; Escherichia-coli ; Negative Chromatography ; Antigen ; Advantages
Funding ProjectNational Natural Sciences Foundation of China[21336010] ; National Natural Sciences Foundation of China[21821005]
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
Funding OrganizationNational Natural Sciences Foundation of China
WOS IDWOS:000453490100009
PublisherELSEVIER SCIENCE BV
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/27590
Collection中国科学院过程工程研究所
Corresponding AuthorZhang, Songping; Su, Zhiguo
Affiliation1.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Li, Zhengjun,Wei, Jiangxue,Yang, Yanli,et al. A two-step heat treatment of cell disruption supernatant enables efficient removal of host cell proteins before chromatographic purification of HBc particles[J]. JOURNAL OF CHROMATOGRAPHY A,2018,1581:71-79.
APA Li, Zhengjun.,Wei, Jiangxue.,Yang, Yanli.,Liu, Lili.,Ma, Guanghui.,...&Su, Zhiguo.(2018).A two-step heat treatment of cell disruption supernatant enables efficient removal of host cell proteins before chromatographic purification of HBc particles.JOURNAL OF CHROMATOGRAPHY A,1581,71-79.
MLA Li, Zhengjun,et al."A two-step heat treatment of cell disruption supernatant enables efficient removal of host cell proteins before chromatographic purification of HBc particles".JOURNAL OF CHROMATOGRAPHY A 1581(2018):71-79.
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