High hydrostatic pressure refolding of highly hydrophobic protein: A case study of recombinant human interferon beta-1b from inclusion bodies

doi:10.1016/j.bej.2021.108055

	High hydrostatic pressure refolding of highly hydrophobic protein: A case study of recombinant human interferon beta-1b from inclusion bodies
	Wang, Qi 2,3; Zhang, Chun 1,3; Li, Zenglan 3; Guo, Fangxia 3; Zhang, Jing 3; Liu, Yongdong 3; Su, Zhiguo 3
刊名	BIOCHEMICAL ENGINEERING JOURNAL
	2021-08-01
卷号	172 页码:9
关键词	High hydrostatic pressure Refolding Hydrophobic protein rhIFN beta-1b Inclusion bodies
ISSN号	1369-703X
DOI	10.1016/j.bej.2021.108055
英文摘要	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.
资助项目	Zhejiang Provincial Natural Science Foundation of China[LQ20H300001] ; National Natural Science Foundation of China[21576267]
WOS关键词	PURIFICATION ; COMPRESSIBILITY ; SOLUBILIZATION ; VOLUME ; MODEL
WOS研究方向	Biotechnology & Applied Microbiology ; Engineering
语种	英语
出版者	ELSEVIER
WOS记录号	WOS:000656859400013
资助机构	Zhejiang Provincial Natural Science Foundation of China ; National Natural Science Foundation of China
内容类型	期刊论文
源URL	[http://ir.ipe.ac.cn/handle/122111/49049]
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.Sichuan Univ, West China Sch Pharm, Minist Educ, Key Lab Drug Targeting & Drug Delivery Syst, Chengdu 610041, 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.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, 1 North Second St, Beijing 100190, Peoples R China
推荐引用方式 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.