Expression, purification, and characterization of recombinant human L-chain ferritin

Protein Expr Purif. 2016 Mar:119:63-8. doi: 10.1016/j.pep.2015.11.018. Epub 2015 Nov 24.

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.

Keywords: Characterization; Expression; Ferritin; Protein cage; Purification; Solubility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoferritins / biosynthesis*
  • Apoferritins / chemistry
  • Apoferritins / isolation & purification
  • Chromatography, Affinity
  • Escherichia coli
  • Gene Expression
  • Humans
  • Nanoparticles / chemistry
  • Particle Size
  • Protein Structure, Secondary

Substances

  • FTL protein, human
  • Apoferritins