Design and characterization of Ni <inf>2+</inf> and Co <inf>2+</inf> decorated Porous Magnetic Silica spheres synthesized by hydrothermal-assisted modified-Stöber method for His-tagged proteins separation

M. Benelmekki, E. Xuriguera, C. Caparros, E. Rodríguez-Carmona, R. Mendoza, J. L. Corchero, S. Lanceros-Mendez, Ll M. Martinez

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27 Citations (Scopus)

Abstract

The complete elimination of enzymes from the reaction mixture and the possibility of its recycling for several rounds result in great benefits, allowing the reduction of the enzyme consumption and their usability in continuous processes. In this work, it is evaluated the capture of a H6-tagged green fluorescence protein (GFP-H6) on porous magnetic spheres using the Co 2+ and Ni 2+ affinity adsorption as a possible cost-effective and up-scaled alternative way for the immobilization of His-tagged proteins. For this purpose, Porous Magnetic Silica (PMS) spheres were synthesized by one-step hydrothermal-assisted modified-Stöber method. The obtained spheres have a homogenous size distribution of 400nm diameter. The γ-Fe 2 O 3 nanoparticles are homogenously distributed in the silica matrix. The obtained PMS spheres have a saturation magnetization of about 10emu/g. Magnetophoresis measurements show a total separation time of 16min at 60T/m. The obtained PMS spheres were successfully and homogenously decorated with Co 2+ and Ni 2+ and then evaluated for the capture of a GFP-H6 protein. The results were compared with the performance of the commercial beads Dynabeads® His-Tag Isolation & Pulldown. © 2011 Elsevier Inc.
Original languageEnglish
Pages (from-to)156-162
JournalJournal of Colloid and Interface Science
Volume365
Issue number1
DOIs
Publication statusPublished - 1 Jan 2012

Keywords

  • His-Tag proteins
  • Magnetic particles
  • Magnetophoresis
  • Metal affinity
  • Surfactant

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