Expression, purification, and tumor uptake of fusion ferritin.
- Author:
Guoyin YAN
1
;
Jun LI
1
;
Ziyang LI
1
Author Information
- Publication Type:Journal Article
- Keywords: ferritin; in vivo imaging; nanoparticles; tumor uptake
- MeSH: Animals; Mice; Recombinant Fusion Proteins/isolation & purification*; Ferritins/metabolism*; Escherichia coli/metabolism*; Melanoma, Experimental/metabolism*; Humans
- From: Chinese Journal of Biotechnology 2025;41(4):1372-1381
- CountryChina
- Language:Chinese
- Abstract: Ferritin is considered as an ideal delivery system due to its precise targeting, reversible self-assembly, high biocompatibility, and easy modification. this study aims to express, purify, and identify three fusion ferritin proteins, and explore their tumor targeting. Three fusion ferritin genes were synthesized and cloned into prokaryotic expression vectors, and the recombinant proteins were purified by affinity chromatography with nickel columns. The fusion ferritin proteins were identified by native polyacrylamide gel electrophoresis (native-PAGE), Western blotting, and circular dichroism. Fluorescein 5-isothiocyanate (FITC) was used to react with fusion ferritin, and confocal laser scanning microscopy was employed to evaluate the tumor targeting of fusion ferritin. The reaction system of sulfo-cyanine7 (Cy7-SE) with fusion ferritin was injected into the tail vein of melanoma mice for in vivo tumor imaging to explore the tumor targeting of fusion ferritin. The results showed that soluble fusion ferritin proteins of about 21 kDa were expressed under the induction by isopropylthio-β-d-galactoside (IPTG), and the recombinant proteins with high purity were obtained. Western blotting showed that the recombinant proteins could be recognized by the corresponding antibodies. The target proteins were identified as multimers with α helixes by native-PAGE and circular dichroism. In vitro and in vivo tumor uptake experiments demonstrated that fusion ferritin was taken up by tumor cells and tumor tissue. This study successfully expressed, purified, and identified fusion ferritin, and verified its tumor uptake in vitro and in vivo, which laid a foundation for the application of ferritin in biomedicine.
