title: Nickel-Salen supported paramagnetic nanoparticles for 6-His-target recombinant protein affinity purification.
abstract: In this research, a simple, efficient, inexpensive, rapid and high yield method for the purification of 6×histidine-tagged recombinant protein was developed. For this purpose, manganese ferrite magnetic nanoparticles (MNPs) were synthesized through a co-precipitation method and then they were conveniently surface-modified with tetraethyl orthosilicate (TEOS) in order to prevent oxidation and form high density of hydroxyl groups. Next, the salen ligand was prepared from condensation reaction of salicylaldehyde and 3-aminopropyl (trimethoxy) silane (APTMS) in 1:1 molar ratio; followed by complexation with Ni(OAc)2.4H2O. Finally, the prepared Ni(II)-salen complex conjugated to silica coated MNPs and MnFe2O4@SiO2@Ni-Salen complex nanoparticles were obtained. The functionalized nanoparticles were spherical with an average diameter around 70nm. The obtained MNPs had a saturation magnetization about 54 emu/g and had super paramagnetic character. These MNPs were used efficiently to enrich recombinant histidine-tagged (His-tagged) protein-A from bacterial cell lysate. In about 45min, highly pure His-tagged recombinant protein was obtained, as judged by SDS-PAGE analysis and silver staining. The amount of target protein in flow through and washing fractions was minimal denoting the high efficiency of purification process. The average capacity of the matrix was found to be high and about 180±15mgg-1(protein/MnFe2O4@SiO2@Ni-Salen complex). Collectively, purification process with MnFe2O4@SiO2@Ni-Salen complex nanoparticles is rapid, efficient, selective and whole purification can be carried out in only a single tube without the need for expensive systems.
authors.affiliation: Department of Organic Chemistry, Faculty of Chemistry, University of Kashan, Kashan, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Rashid
authors.affiliation: Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
authors.forename: Ramin
authors.initials: R
authors.lastname: Ghahremanzadeh
authors.affiliation: Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Nanotechnology Research Center, Faculty of Pharamcy, Tehran University of Medical Sciences (TUMS), Tehran, Iran.
authors.forename: Mohammad-Reza
authors.initials: MR
authors.lastname: Nejadmoghaddam
authors.affiliation: Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
authors.forename: Mahboobeh
authors.initials: M
authors.lastname: Nazari
authors.affiliation: Department of Immunology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Mohammad-Reza
authors.initials: MR
authors.lastname: Shokri
authors.affiliation: Department of Organic Chemistry, Faculty of Chemistry, University of Kashan, Kashan, Iran. Electronic address: naeimi@kashanu.ac.ir.
authors.forename: Hossein
authors.initials: H
authors.lastname: Naeimi
authors.affiliation: Department of Immunology, School of Public Health, Tehran University of Medical Science, Tehran, Iran; Immunology Research Center, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Amir-Hassan
authors.initials: AH
authors.lastname: Zarnani
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D005029:Ethylenediamines; D058185:Magnetite Nanoparticles; D009532:Nickel; D011994:Recombinant Proteins / Q000302:isolation & purification*; D012821:Silanes; D012822:Silicon Dioxide
chemical_list: D005029:Ethylenediamines; D058185:Magnetite Nanoparticles; D011994:Recombinant Proteins; D012821:Silanes; C040733:tetraethoxysilane; D012822:Silicon Dioxide; D009532:Nickel; C011452:disalicylaldehyde ethylenediamine
keywords: Histidine-tagged protein; Immobilized metal affinity chromatography (IMAC); Magnetic nanoparticles; Protein purification; Salen-type ligands