Title
Conversion of recombinant human ferritin light chain inclusion bodies into uniform nanoparticles in Escherichia coli for facile production
Author
Xiaotong Song, Yongxiang Zheng, Yongdong Liu, Huan Meng, Rong Yu, Chun Zhang
Year
2022
Journal
Engineering
Abstract
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.
Instrument
J-810
Keywords
Prokaryotic expression, rhFTL, structure