CAS OpenIR
Development of purification process for dual-function recombinant human heavy-chain ferritin by the investigation of genetic modification impact on conformation
Yin, Shuang1; Zhang, Bingyang1; Lin, Jianying2; Liu, Yongdong3; Su, Zhiguo3; Bi, Jingxiu1
2021-06-19
Source PublicationENGINEERING IN LIFE SCIENCES
ISSN1618-0240
Pages13
AbstractFerritin is a promising drug delivery platform and has been functionalized through genetic modifications. This work has designed and expressed a dual-functional engineered human heavy-chain ferritin (HFn) with the inserted functional peptide PAS and RGDK to extend half-life and improve tumor targeted drug delivery. A facile and cost-effective two-step purification pathway for recombinant HFn was developed. The genetic modification was found to affect HFn conformation, and therefore varied the purification performance. Heat-acid precipitation followed by butyl fast flow hydrophobic interaction chromatography (HIC) has been developed to purify HFn and modified HFns. Nucleic acid removal reached above 99.8% for HFn and modified HFns. However, HFn purity reached above 95% and recovery yield (overall) above 90%, compared with modified HFns purity above 82% and recovery yield (overall) above 58%. It is interesting to find that the inserted functional peptides significantly changed the molecule conformation, where a putative turnover of the E-helix with the inserted functional peptides formed a "flop" conformation, in contrast with the "flip" conformation of HFn. It could be the cause of fragile stability of modified HFns, and therefore less tolerant to heat and acid condition, observed by the lower recovery yield in heat-acid precipitation.
Keywordferritin E-helix turnover ferritin genetic modification human heavy-chain ferritin protein characterization protein purification
DOI10.1002/elsc.202000105
Language英语
WOS KeywordHALF-LIFE ; H-CHAIN ; PROTEINS ; DELIVERY ; BINDING ; NANOPARTICLES ; NANOCARRIERS ; PLATFORM
Funding ProjectUniversity of Adelaide ; Institute of Process Engineering, Chinese Academy of Sciences ; National Natural Science Foundation of China[21576267] ; Beijing Natural Science Foundation[2162041]
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
Funding OrganizationUniversity of Adelaide ; Institute of Process Engineering, Chinese Academy of Sciences ; National Natural Science Foundation of China ; Beijing Natural Science Foundation
WOS IDWOS:000663354600001
PublisherWILEY
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/49199
Collection中国科学院过程工程研究所
Corresponding AuthorBi, Jingxiu
Affiliation1.Univ Adelaide, Sch Chem Engn & Adv Mat, Fac Engn Comp & Math Sci, North Terrace, Adelaide, SA 5005, Australia
2.Taiyuan Univ Technol, Coll Biomed Engn, Taiyuan, Peoples R China
3.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing, Peoples R China
Recommended Citation
GB/T 7714
Yin, Shuang,Zhang, Bingyang,Lin, Jianying,et al. Development of purification process for dual-function recombinant human heavy-chain ferritin by the investigation of genetic modification impact on conformation[J]. ENGINEERING IN LIFE SCIENCES,2021:13.
APA Yin, Shuang,Zhang, Bingyang,Lin, Jianying,Liu, Yongdong,Su, Zhiguo,&Bi, Jingxiu.(2021).Development of purification process for dual-function recombinant human heavy-chain ferritin by the investigation of genetic modification impact on conformation.ENGINEERING IN LIFE SCIENCES,13.
MLA Yin, Shuang,et al."Development of purification process for dual-function recombinant human heavy-chain ferritin by the investigation of genetic modification impact on conformation".ENGINEERING IN LIFE SCIENCES (2021):13.
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