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

摘要

The present study is aimed at the synthesis of MnFe _(2) O _(4) @SiO _(2) @NH _(2) @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 MnFe _(2) O _(4) . Synthesized MnFe _(2) O _(4) @SiO _(2) was then properly NH _(2) -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 MnFe _(2) O _(4) @SiO _(2) . The stable surface functionalized nanoparticles were further linked with Ni ~(2+) and used in powder form for efficient purification of 6×His-tagged proteins from the mixture of lysed cells. MnFe _(2) O _(4) @SiO _(2) @NH _(2) @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.