Polymer brush-modified magnetic nanoparticles for His-tagged protein purification

Langmuir. 2011 Mar 15;27(6):3106-12. doi: 10.1021/la1050404. Epub 2011 Feb 21.

Abstract

Growth of poly(2-hydroxyethyl methacrylate) brushes on magnetic nanoparticles and subsequent brush functionalization with nitrilotriacetate-Ni(2+) yield magnetic beads that selectively capture polyhistidine-tagged (His-tagged) protein directly from cell extracts. Transmission electron microscopy, Fourier transform infrared (FT-IR) spectroscopy, thermogravimetric analysis, and magnetization measurements confirm and quantify the formation of the brushes on magnetic particles, and multilayer protein adsorption to these brushes results in binding capacities (220 mg BSA/g of beads and 245 mg His-tagged ubiquitin/g of beads) that are an order of magnitude greater than those of commercial magnetic beads. Moreover, the functionalized beads selectively capture His-tagged protein within 5 min. The high binding capacity and protein purity along with efficient protein capture in a short incubation time make brush-modified particles attractive for purification of recombinant proteins.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Histidine / chemistry*
  • Magnetite Nanoparticles / chemistry*
  • Molecular Structure
  • Particle Size
  • Polyhydroxyethyl Methacrylate / chemical synthesis
  • Polyhydroxyethyl Methacrylate / chemistry*
  • Surface Properties
  • Ubiquitin / chemistry
  • Ubiquitin / isolation & purification*

Substances

  • Magnetite Nanoparticles
  • Ubiquitin
  • Polyhydroxyethyl Methacrylate
  • Histidine