Synthesis of silica nanospheres with Ni2+-iminodiacetic acid for protein separation
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Silica (SiO2) nanospheres (NSs) with immobilized metal ligands have been prepared for the affinity separation of proteins. First, SiO2 NSs were prepared by controlled hydrolysis of tetraethoxysilane in a basic aqueous-ethanol solution. Then through reaction of iminodiacetic acid (IDA) with 3-glycidoxypropyltrimethoxysilane and immobilization of them onto the surfaces of above SiO2 NSs, novel affinity adsorbents with IDA chelating groups were obtained. After chelating Ni2+ ions, the SiO2–IDA–Ni2+ NSs were applied to separate his-tagged proteins directly from the mixture of lysed cells. The SiO2–IDA–Ni2+ NSs present negligible nonspecific protein adsorption and high protein binding ability (28.3 mg/g).
KeywordsSilica nanospheres Surface modification Protein separation His-tagged protein
Financial support of this work from National Natural Science Foundation of China (21271062) and Construction Fund of Henan province and Department of Education (SBGJ090515) is gratefully acknowledged.
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