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High hydrostatic pressure refolding of highly hydrophobic protein: A case study of recombinant human interferon beta-1b from inclusion bodies
Wang, Qi1,2; Zhang, Chun1,3; Li, Zenglan1; Guo, Fangxia1; Zhang, Jing1; Liu, Yongdong1; Su, Zhiguo1
2021-08-01
Source PublicationBIOCHEMICAL ENGINEERING JOURNAL
ISSN1369-703X
Volume172Pages:9
AbstractRecombinant human interferon beta-lb (rhIFN beta-lb) represents a highly hydrophobic protein and is difficult to refold from inclusion bodies (IBs) using conventional methods. In this paper, we report a strategy of high hydrostatic pressure (HHP) combining with additives for facile refolding of rhIFN beta-lb IBs: 1) refining the rhIFN beta-lb IBs with 8 M urea supplemented with 1% Triton X-100 to reduce the amount of impurities; 2) adding 5 mM DTT to enable 100 % mass yield of rhIFN beta-lb IBs; 3) addition of zwittergent 3-14 could significantly enhance the initial refolding concentration while did not disturb the following purification; 4) maintaining a pressure of 320 MPa could ensure considerable refolding efficacy of rhIFN beta-lb IBs. As a result, refolding of rhIFN beta-lb IBs with optimal additives was under 320 MPa for 16 h, almost 80 % refolding yield was achieved even with an initial rhIFN beta-lb IBs suspension of 4 mg/mL. After purification by SP-Sepharose fast flow (SP FF) and Butyl-S fast flow (Butyl-S FF), an overall yield of 37.8 % of rhIFN beta-lb was achieved. The resulted rhIFN beta-lb showed similar structures and comparable in vitro bioactivity to rhIFN beta-lb standard. The whole process featured integration of solubilization/refolding with high initial refolding concentration and considerable yield, indicating HHP refolding is a promising strategy for production of highly hydrophobic proteins from IBs.
KeywordHigh hydrostatic pressure Refolding Hydrophobic protein rhIFN beta-1b Inclusion bodies
DOI10.1016/j.bej.2021.108055
Language英语
WOS KeywordPURIFICATION ; COMPRESSIBILITY ; SOLUBILIZATION ; VOLUME ; MODEL
Funding ProjectZhejiang Provincial Natural Science Foundation of China[LQ20H300001] ; National Natural Science Foundation of China[21576267]
WOS Research AreaBiotechnology & Applied Microbiology ; Engineering
WOS SubjectBiotechnology & Applied Microbiology ; Engineering, Chemical
Funding OrganizationZhejiang Provincial Natural Science Foundation of China ; National Natural Science Foundation of China
WOS IDWOS:000656859400013
PublisherELSEVIER
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/49050
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, State Key Lab Biochem Engn, 1 North Second St, Beijing 100190, Peoples R China
2.Zhejiang Univ, Coll Med, Affiliated Hosp 1,Ctr Clin Lab, Key Lab Clin In Vitro Diagnost Tech Zhejiang Prov, Hangzhou 310003, Zhejiang, Peoples R China
3.Sichuan Univ, West China Sch Pharm, Minist Educ, Key Lab Drug Targeting & Drug Delivery Syst, Chengdu 610041, Peoples R China
Recommended Citation
GB/T 7714
Wang, Qi,Zhang, Chun,Li, Zenglan,et al. High hydrostatic pressure refolding of highly hydrophobic protein: A case study of recombinant human interferon beta-1b from inclusion bodies[J]. BIOCHEMICAL ENGINEERING JOURNAL,2021,172:9.
APA Wang, Qi.,Zhang, Chun.,Li, Zenglan.,Guo, Fangxia.,Zhang, Jing.,...&Su, Zhiguo.(2021).High hydrostatic pressure refolding of highly hydrophobic protein: A case study of recombinant human interferon beta-1b from inclusion bodies.BIOCHEMICAL ENGINEERING JOURNAL,172,9.
MLA Wang, Qi,et al."High hydrostatic pressure refolding of highly hydrophobic protein: A case study of recombinant human interferon beta-1b from inclusion bodies".BIOCHEMICAL ENGINEERING JOURNAL 172(2021):9.
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