Journal
RSC ADVANCES
Volume 6, Issue 43, Pages 36840-36848Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c5ra25949e
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Funding
- Iran National Science Foundation (INSF) [92027471]
- University of Kashan [159148/34]
- Avicenna Research Institute
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The present study is aimed at the synthesis of MnFe2O4@SiO2@NH2@2AB-Ni, as a highly efficient and novel affinity adsorbent, for specific purification of 6 x histidine-tagged recombinant proteins. The new immobilized metal ion affinity adsorbent was fabricated following co-precipitation synthesis of superparamagnetic manganese ferrite nanoparticles. Subsequently, tetraethyl orthosilicate (TEOS) was added in weak basic conditions (pH similar to 9) to prevent oxidation and increase the density of -OH groups on the surface of MnFe2O4. Synthesized MnFe2O4@SiO2 was then properly NH2-functionalized with 3-aminopropyl-trimethoxylsilane (APTMS) as anchor molecules. Manganese ferrite nanoparticles were converted to bidentate ligands through a reaction between isatoic anhydride and amino-functionalized MnFe2O4@SiO2. The stable surface functionalized nanoparticles were further linked with Ni2+ and used in powder form for efficient purification of 6 x His-tagged proteins from the mixture of lysed cells. MnFe2O4@SiO2@NH2@2AB-Ni nanoparticles exhibited excellent performance in the separation of 6x histidine-tagged recombinant protein-A from cell lysate, with a binding capacity of about 220 mg g(-1). Indeed, the synthesized magnetic nanoparticles presented negligible nonspecific protein adsorption.
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