Conversion of recombinant human ferritin light chain inclusion bodies into uniform nanoparticles in Escherichia coli for facile production

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 i...

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Published inEngineering in life sciences Vol. 22; no. 6; pp. 453 - 463
Main Authors Song, Xiaotong, Zheng, Yongxiang, Liu, Yongdong, Meng, Huan, Yu, Rong, Zhang, Chun
Format Journal Article
LanguageEnglish
Published Germany John Wiley & Sons, Inc 01.06.2022
John Wiley and Sons Inc
Wiley-VCH
Subjects
Bacteria
Capto Butyl
Chains
Chromatography
Cloning
E coli
E. coli soluble expression
Escherichia coli
Ferritin
Gene expression
Glycerol
Impurities
Inclusion bodies
Ionic strength
Light
Mass spectrometry
Mass spectroscopy
Nanoparticles
pBV220 plasmid
precipitation procedure
Proteins
recombinant human ferritin light chain
United Kingdom > UK
United States > US
China
Japan
E. coli soluble expression
pBV220 plasmid
precipitation procedure
Capto Butyl
recombinant human ferritin light chain
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Summary: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.
Bibliography:Xiaotong Song and Yongxiang Zheng contributed equally to this work, were the co‐first authors.
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ISSN:1618-0240
1618-2863
DOI:10.1002/elsc.202100164