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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 Publication | BIOCHEMICAL ENGINEERING JOURNAL
 |
| ISSN | 1369-703X |
| Volume | 172Pages:9 |
| Abstract | Recombinant 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. |
| Keyword | High hydrostatic pressure Refolding Hydrophobic protein rhIFN beta-1b Inclusion bodies |
| DOI | 10.1016/j.bej.2021.108055 |
| Language | 英语 |
| WOS Keyword | PURIFICATION ; COMPRESSIBILITY ; SOLUBILIZATION ; VOLUME ; MODEL |
| Funding Project | Zhejiang Provincial Natural Science Foundation of China[LQ20H300001] ; National Natural Science Foundation of China[21576267] |
| WOS Research Area | Biotechnology & Applied Microbiology ; Engineering |
| WOS Subject | Biotechnology & Applied Microbiology ; Engineering, Chemical |
| Funding Organization | Zhejiang Provincial Natural Science Foundation of China ; National Natural Science Foundation of China |
| WOS ID | WOS:000656859400013 |
| Publisher | ELSEVIER |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.ipe.ac.cn/handle/122111/49050 |
| Collection | 中国科学院过程工程研究所 |
| Corresponding Author | Liu, Yongdong; Su, Zhiguo |
| Affiliation | 1.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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