Fast and Highly Efficient Purification of 6×Histidine-Tagged Recombinant Proteins by Ni-Decorated Mnfe2o4@Sio2@Nh2@2Ab As Novel and Efficient Affinity Adsorbent Magnetic Nanoparticles Publisher



Rashid Z¹ ; Naeimi H¹ ; Zarnani AH^{2, 3} ; Nazari M⁴ ; Nejadmoghaddam MR^{4, 5} ; Ghahremanzadeh R⁴ Show Affiliations
Source: RSC Advances Published:2016

Abstract

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×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 ∼ 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×His-tagged proteins from the mixture of lysed cells. MnFe2O4@SiO2@NH2@2AB-Ni nanoparticles exhibited excellent performance in the separation of 6×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. © The Royal Society of Chemistry 2016.