Conversion of recombinant human ferritin light chain inclusion bodies into uniform nanoparticles in Escherichia coli for facile production
文献类型:期刊论文
作者 | Song, Xiaotong1,2; Zheng, Yongxiang1,2; Liu, Yongdong3; Meng, Huan1,2; Yu, Rong1,2; Zhang, Chun1,2 |
刊名 | ENGINEERING IN LIFE SCIENCES
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出版日期 | 2022-04-23 |
页码 | 11 |
关键词 | Capto Butyl E coli soluble expression pBV220 plasmid precipitation procedure recombinant human ferritin light chain |
ISSN号 | 1618-0240 |
DOI | 10.1002/elsc.202100164 |
英文摘要 | Prokaryotic expression systems are widely used to produce many types of biologics because of their extreme conveniences and unmatchable cost. However, production of recombinant human ferritin light chain (rhFTL) protein is largely restrained because its expression in Escherichia coli tends to form inclusion bodies (IBs). In this study, a prokaryotic expression vector (FTL-pBV220) harboring the rhFTL gene was constructed using a pBV220 plasmid. The tag-free rhFTL was highly expressed and almost entirely converted to soluble form, and thus the rhFTL was successfully self-assembled into uniform nanoparticles in E. coli. To establish a simplified downstream process, a precipitation procedure based on the optimized incubation temperature, pH condition, and ionic strength was developed to remove impurities from the crude lysate supernatant. The rhFTL retained in the clarified supernatant was subsequently purified in a single step using Capto Butyl column resulting in a considerable recovery and high purity. The purified rhFTL was characterized and verified by mass spectrometry and spectral and morphological analyses. The results revealed that rhFTL exhibited highly ordered and fairly compact structures and the spherical structures were preserved. |
WOS关键词 | PURIFICATION |
资助项目 | National Natural Science Foundation of China[81773623] ; National Natural Science Foundation of China[81803420] ; National Natural Science Foundation of China[81973226] ; National Natural Science Foundation of China[82003685] ; Sichuan Science and Technology Program[2021YJ0187] ; Fundamental Research Funds for the Central Universities[YJ201926] ; 111-project[B18035] |
WOS研究方向 | Biotechnology & Applied Microbiology |
语种 | 英语 |
WOS记录号 | WOS:000785911000001 |
出版者 | WILEY |
资助机构 | National Natural Science Foundation of China ; Sichuan Science and Technology Program ; Fundamental Research Funds for the Central Universities ; 111-project |
源URL | [http://ir.ipe.ac.cn/handle/122111/52722] ![]() |
专题 | 中国科学院过程工程研究所 |
通讯作者 | Zhang, Chun |
作者单位 | 1.Sichuan Univ, West China Sch Pharm, Sichuan Engn Lab Plant Sourced Drug,Educ Minist, Dept Biopharmaceut,Key Lab Drug Targeting & Drug, 17,Sect 3,Southern Renmin Rd, Chengdu 610041, Peoples R China 2.Sichuan Univ, West China Sch Pharm, Sichuan Res Ctr Drug Precis Ind Technol, 17,Sect 3,Southern Renmin Rd, Chengdu 610041, Peoples R China 3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing, Peoples R China |
推荐引用方式 GB/T 7714 | Song, Xiaotong,Zheng, Yongxiang,Liu, Yongdong,et al. Conversion of recombinant human ferritin light chain inclusion bodies into uniform nanoparticles in Escherichia coli for facile production[J]. ENGINEERING IN LIFE SCIENCES,2022:11. |
APA | Song, Xiaotong,Zheng, Yongxiang,Liu, Yongdong,Meng, Huan,Yu, Rong,&Zhang, Chun.(2022).Conversion of recombinant human ferritin light chain inclusion bodies into uniform nanoparticles in Escherichia coli for facile production.ENGINEERING IN LIFE SCIENCES,11. |
MLA | Song, Xiaotong,et al."Conversion of recombinant human ferritin light chain inclusion bodies into uniform nanoparticles in Escherichia coli for facile production".ENGINEERING IN LIFE SCIENCES (2022):11. |
入库方式: OAI收割
来源:过程工程研究所
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