Ying Yang y.yang@keele.ac.uk
Synthesis of magnetic nanoparticles with an IDA or TED modified surface for purification and immobilization of poly-histidine tagged proteins
Yang
Authors
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.
Acceptance Date | Mar 2, 2020 |
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Publication Date | Mar 20, 2020 |
Journal | RSC Advances |
Print ISSN | 2046-2069 |
Publisher | Royal Society of Chemistry |
Pages | 11524 - 11534 |
DOI | https://doi.org/10.1039/C9RA10473A |
Publisher URL | https://doi.org/10.1039/C9RA10473A |
Files
c9ra10473a.pdf
(1.6 Mb)
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Publisher Licence URL
https://creativecommons.org/licenses/by-nc/4.0/
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