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Preparation of core–shell structure Fe 3 O 4 @SiO 2 superparamagnetic microspheres immoblized with iminodiacetic acid as immobilized metal ion affinity adsorbents for His‐tag protein purification
Author(s) -
Ni Qian,
Chen Bing,
Dong Shaohua,
Tian Lei,
Bai Quan
Publication year - 2016
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3584
Subject(s) - iminodiacetic acid , chemistry , adsorption , superparamagnetism , metal ions in aqueous solution , chelation , chromatography , metal , magnetic nanoparticles , elution , lysozyme , protein purification , selective adsorption , ion exchange , nuclear chemistry , inorganic chemistry , ion , magnetization , nanotechnology , materials science , nanoparticle , organic chemistry , magnetic field , biochemistry , physics , quantum mechanics
The core–shell structure Fe 3 O 4 /SiO 2 magnetic microspheres were prepared by a sol–gel method, and immobiled with iminodiacetic acid (IDA) as metal ion affinity ligands for protein adsorption. The size, morphology, magnetic properties and surface modification of magnetic silica nanospheres were characterized by various modern analytical instruments. It was shown that the magnetic silica nanospheres exhibited superparamagnetism with saturation magnetization values of up to 58.1 emu/g. Three divalent metal ions, Cu 2+ , Ni 2+ and Zn 2+ , were chelated on the Fe 3 O 4 @SiO 2 –IDA magnetic microspheres to adsorb lysozyme. The results indicated that Ni 2+ ‐chelating magnetic microspheres had the maximum adsorption capacity for lysozyme of 51.0 mg/g, adsorption equilibrium could be achieved within 60 min and the adsorbed protein could be easily eluted. Furthermore, the synthesized Fe 3 O 4 @SiO 2 –IDA–Ni 2+ magnetic microspheres were successfully applied for selective enrichment lysozyme from egg white and His‐tag recombinant Homer 1a from the inclusion extraction expressed in Escherichia coli . The result indicated that the magnetic microspheres showed unique characteristics of high selective separation behavior of protein mixture, low nonspecific adsorption, and easy handling. This demonstrates that the magnetic silica microspheres can be used efficiently in protein separation or purification and show great potential in the pretreatment of the biological sample. Copyright © 2015 John Wiley & Sons, Ltd.