title: Super magnetic nanoparticles NiFe2O4, coated with aluminum-nickel oxide sol-gel lattices to safe, sensitive and selective purification of his-tagged proteins.
abstract: Super magnetic nanoparticle NiFe2O4 with high magnetization, physical and chemical stability was introduced as a core particle which exhibits high thermal stability (>97%) during the harsh coating process. Instead of multi-stage process for coating, the magnetic nanoparticles was mineralized via one step coating by a cheap, safe, stable and recyclable alumina sol-gel lattice (from bohemite source) saturated by nickel ions. The TEM, SEM, VSM and XRD imaging and BET analysis confirmed the structural potential of NiFe2O4@NiAl2O4 core-shell magnetic nanoparticles for selective and sensitive purification of His-tagged protein, in one step. The functionality and validity of the nickel magnetic nanoparticles were attested by purification of three different bioactive His-tagged recombinant fusion proteins including hIGF-1, GM-CSF and bFGF. The bonding capacity of the nickel magnetics nanoparticles was studied by Bradford assay and was equal to 250 ± 84 μg Protein/mg MNP base on protein size. Since the metal ion leakage is the most toxicity source for purification by nickel magnetic nanoparticles, therefor the nickel leakage in purified final protein was determined by atomic absorption spectroscopy and biological activity of final purified protein was confirmed in comparison with reference. Also, in vitro cytotoxicity of nickel magnetic nanoparticles and trace metal ions were investigated by MTS assay analysis. The results confirmed that the synthesized nickel magnetic nanoparticles did not show metal ion toxicity and not affected on protein folding.
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran. Electronic address: mirahmadi_zare@royaninstitute.org.
authors.forename: Seyede Zohreh
authors.initials: SZ
authors.lastname: Mirahmadi-Zare
authors.affiliation: Nanotechnology and Advanced Materials Institute, Isfahan University of Technology, Isfahan, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Allafchian
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Aboutalebi
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Pendar
authors.initials: P
authors.lastname: Shojaei
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Yahya
authors.initials: Y
authors.lastname: Khazaie
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Kianoush
authors.initials: K
authors.lastname: Dormiani
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Liana
authors.initials: L
authors.lastname: Lachinani
authors.affiliation: Department of Molecular Biotechnology, Cell Science Research Center, Royan Institute for Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Mohammad-Hossein
authors.initials: MH
authors.lastname: Nasr-Esfahani
mesh_terms: D000535:Aluminum / Q000737:chemistry; D005290:Ferric Compounds / Q000737:chemistry*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000737:chemistry / Q000302:isolation & purification; D006639:Histidine / Q000737:chemistry; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000737:chemistry / Q000302:isolation & purification; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D009532:Nickel / Q000737:chemistry*; D044367:Phase Transition; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D005290:Ferric Compounds; D058185:Magnetite Nanoparticles; D011993:Recombinant Fusion Proteins; C550717:nickel ferrite; D006639:Histidine; D007334:Insulin-Like Growth Factor I; D009532:Nickel; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; C028007:nickel monoxide; D000535:Aluminum
keywords: Alumina solgel coating; Bioactivity; Histidine tagged protein; Nickel magnetic nanoparticles; Protein purification