title: Expression, purification, and characterization of recombinant human L-chain ferritin.
abstract: Ferritins form nanocage architectures and demonstrate their potential to serve as functional nanomaterials with potential applications in medical imaging and therapy. In our study, the cDNA of human L-chain ferritin was cloned into plasmid pET-28a for its overexpression in Escherichia coli. However, the recombinant human L-chain ferritin (rLF) was prone to form inclusion bodies. Molecular chaperones were co-expressed with rLF to facilitate its correct folding. Our results showed that the solubility of rLF was increased about 3-fold in the presence of molecular chaperones, including GroEL, GroES and trigger factor. Taking advantage of its N-terminal His-tag, rLF was then purified with Ni-affinity chromatography. With a yield of 10 mg/L from bacterial culture, the purified rLF was analyzed by circular dichroism spectrometry for its secondary structure. Furthermore, the rLF nanocages were characterized using dynamic light scattering and transmission electron microscopy.
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China.
authors.forename: Wenyan
authors.initials: W
authors.lastname: Zou
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China.
authors.forename: Xi
authors.initials: X
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China.
authors.forename: Dianhua
authors.initials: D
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China.
authors.forename: Jiahuang
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China. Electronic address: jilina@nju.edu.cn.
authors.forename: Lina
authors.initials: L
authors.lastname: Ji
authors.affiliation: State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing 210046, Jiangsu, PR China. Electronic address: huazc@nju.edu.cn.
authors.forename: Zichun
authors.initials: Z
authors.lastname: Hua
mesh_terms: D001052:Apoferritins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004926:Escherichia coli; D015870:Gene Expression; D006801:Humans; D053758:Nanoparticles / Q000737:chemistry; D010316:Particle Size; D017433:Protein Structure, Secondary
chemical_list: C507433:FTL protein, human; D001052:Apoferritins
keywords: Characterization; Expression; Ferritin; Protein cage; Purification; Solubility