title: Expression, purification, and characterization of recombinant human H-chain ferritin.
abstract: Based on their nanocage architectures, ferritins show their potential applications in medical imaging and therapeutic delivery systems. However, the recombinant human H-chain ferritin (rHF) is prone to form inclusion bodies in Escherichia coli. In our study, the cDNA of rHF was cloned into plasmid pET28a under the control of a T7 promoter. Molecular chaperones, including GroES, GroEL, and trigger factor, were coexpressed with rHF to facilitate its correct folding. The results showed that the solubility of rHF was increased more than threefold with the help of molecular chaperones. Taking advantages of its N-terminal His-tag, rHF was then purified with Ni-affinity chromatography. With a yield of 15 mg/L from bacterial culture, the purified rHF was analyzed by circular dichroism spectrometry for its secondary structure. Moreover, the rHF nanocages were characterized by transmission electron microscopy and dynamic light scattering. Our results indicate that rHF is able to self-assemble into nanocages with a narrow size distribution.
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Wenyan
authors: W
authors: Zou
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Xiaoyu
authors: X
authors: Liu
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Dianhua
authors: D
authors: Chen
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Jie
authors: J
authors: Wang
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Xi
authors: X
authors: Zhao
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Jiahuang
authors: J
authors: Li
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Lina
authors: L
authors: Ji
authors: a State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences , Nanjing University , Nanjing , Jiangsu , China.
authors: Zichun
authors: Z
authors: Hua
mesh_terms: D001052:Apoferritins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000648:ultrastructure; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D017510:Protein Folding; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000648:ultrastructure; D012995:Solubility
chemical_list: D011994:Recombinant Proteins; D001052:Apoferritins
keywords: Characterization; expression; ferritin; protein cage; purification; solubility