Layer by layer coating of NH2-silicate/polycarboxylic acid polymer saturated by Ni2+ onto the super magnetic NiFe2O4 nanoparticles for sensitive and bio-valuable separation of His-tagged proteins.

Author(s) Mirahmadi-Zare, S.Zohreh; Aboutalebi, F.; Allafchian, M.; Pirjamali, L.; Nasr-Esfahani, M.H.
Journal Protein Expr Purif
Date Published 2018 Mar
Abstract

Magnetic nanoparticles NiFe2O4 was synthesized and covered in the silicate lattice of (3-Aminopropyl) triethoxysilane (APS) by the sol-gel process. Subsequently, the EDTA-dianhydride was attached to the amino surface of magnetic nanoparticles (MNPs) during the nucleophilic attack. This polycarboxylic layer trapped the high level of nickel ions for selective bonding to the His-tagged recombinant protein. The surface of MNPs was investigated by TEM, XRD, SEM (EDSA), VSM, BET, FT-IR and zeta potential analysis which characterized the size, chemical lattice, morphology, magnetic strength, specific surface area, functional groups and charge of the surface of nanoparticles. The performance and validity of the nanoparticles were studied by the purification of His-tagged green fluorescence protein (His-GFP). Also, the safety of proposed Ni-MNPs in the purification procedure of His-tagged proteins for pharmaceutical applications was proved by the determination of the nickel leakage level in the purified final protein using atomic absorption spectroscopy. In vitro cytotoxicity of Ni-MNPs and trace metal ions was investigated by the MTS assay technique. In addition, the comparison of biological activity in purified protein (GM-CSF) and commercial sample did not show any toxic effect.

DOI 10.1016/j.pep.2017.10.015
ISSN 1096-0279
Citation Mirahmadi-Zare SZ, Aboutalebi F, Allafchian M, Pirjamali L, Nasr-Esfahani M-. Layer by layer coating of NH2-silicate/polycarboxylic acid polymer saturated by Ni2+ onto the super magnetic NiFe2O4 nanoparticles for sensitive and bio-valuable separation of His-tagged proteins. Protein Expr Purif. 2018;143:71-76.

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