Purification of 6×His-tagged phycobiliprotein using zinc-decorated silica-coated magnetic nanoparticles

J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Apr 15;879(13-14):993-7. doi: 10.1016/j.jchromb.2011.03.014. Epub 2011 Mar 23.

Abstract

Zinc-decorated silica-coated magnetic nanoparticles (ZnSiMNPs) were prepared by adsorbing zinc onto colloidal silica. These nanoparticles were used for the rapid purification of 6×His-tagged recombinant phycobiliprotein. The surface changes in the magnetic nanoparticles after zinc adsorption were characterized by transmission electron microscopy. The adsorption of the 6×His-tagged phycobiliprotein onto ZnSiMNPs in 10 mM PBS at 25°C was found to follow the Langmuir isotherm. ZnSiMNPs could be used to extract 6×His-tagged phycobiliprotein from lysate to single-band purity, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. No spectral variation was observed in the purified phycobiliprotein. Thus, ZnSiMNPs served as a useful tool for the magnetic separation and delivery of the 6×His-tagged phycobiliprotein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Chromatography, Affinity / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Histidine / chemistry
  • Histidine / isolation & purification*
  • Magnetite Nanoparticles / chemistry*
  • Microscopy, Electron, Transmission
  • Oligopeptides / chemistry
  • Oligopeptides / isolation & purification*
  • Particle Size
  • Phycobiliproteins / chemistry
  • Phycobiliproteins / isolation & purification*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / isolation & purification*
  • Silicon Dioxide / chemistry
  • Zinc / chemistry

Substances

  • His-His-His-His-His-His
  • Magnetite Nanoparticles
  • Oligopeptides
  • Phycobiliproteins
  • Recombinant Fusion Proteins
  • Histidine
  • Silicon Dioxide
  • Zinc