Zeng, K, Sun, E-J, Liu, Z-W, Guo, J, Yuan, C, Yang, Y ORCID: https://orcid.org/0000-0002-4292-627X and Xie, H (2020) Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins. RSC Advances, 10. 11524 - 11534.

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Abstract

Magnetic nanoparticles (MNPs) chelating with metal ions can specifically interact with poly-histidine peptides and facilitate immobilization and purification of proteins with poly-histidine tags. Fabrication of MNPs is generally complicated and time consuming. In this paper, we report the preparation of Ni(ii) ion chelated MNPs (Ni-MNPs) in two stages for protein immobilization and purification. In the first stage, organic ligands including pentadentate tris (carboxymethyl) ethylenediamine (TED) and tridentate iminodiacetic acid (IDA) and inorganic Fe3O4-SiO2 MNPs were synthesized separately. In the next stage, ligands were grafted to the surface of MNPs and MNPs with a TED or IDA modified surface were acquired, followed by chelating with Ni(ii) ions. The Ni(ii) ion chelated forms of MNPs (Ni-MNPs) were characterized including morphology, surface charge, structure, size distribution and magnetic response. Taking a his-tagged glycoside hydrolase DspB (Dispersin B) as the protein representative, specific interactions were confirmed between DspB and Ni-MNPs. Purification of his-tagged DspB was achieved with Ni-MNPs that exhibited better performance in terms of purity and activity of DspB than commercial Ni-NTA. Ni-MNPs as enzyme carriers for DspB also exhibited good compatibility and reasonable reusability as well as improved performance in various conditions.

Item Type: Article
Subjects: Q Science > QD Chemistry
Divisions: Faculty of Medicine and Health Sciences > Institute for Science and Technology in Medicine
Depositing User: Symplectic
Date Deposited: 06 Aug 2020 12:56
Last Modified: 06 Aug 2020 12:56
URI: https://eprints.keele.ac.uk/id/eprint/8511

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