Fe3O4@PAM@NTA-Ni2+ Magnetic Composite Nanoparticles for Highly Specific Separation of His-tagged Proteins |
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| Authors: | Huiling Guo Honghao Sun Zhengding Su Shuchao Hu Xiaotao Wang |
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| Affiliation: | 1.School of Food and Pharmaceutical Engineering, Hubei Provincial Cooperative Innovation Center of Industrial Fermentation,Hubei University of Technology,Wuhan,China;2.School of Biological Engineering, Hubei Provincial Cooperative Innovation Center of Industrial Fermentation,Hubei University of Technology,Wuhan,China;3.Shool of Materials Science and Engineering,Hubei University of Technology,Wuhan,China |
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| Abstract: | ![]()
A facile approach has been developed to synthesize Fe3O4@PAM (polyacrylamide) nanoparticles (NPs) with carboxyl groups on the surfaces by copolymerization with acrylamide and acrylic acid in Fe3O4 NPs aqueous suspension. Nitrilotriacetic acid (NTA) was conjugated to the magnetic NPs via well-known carboniimide chemistry using EDC and NHS. The Ni2+ ions loaded on the surface of NPs provide abundant docking sites for immobilization of His-tagged green fluorescent proteins (His-tagged GFP). The high magnetic property of Fe3O4@PAM@NTA-Ni2+ allows an easy separation of the NPs from solution under an external magnetic field, with high His-tagged protein binding capacity (42 μg protein/mg of NPs). The NPs can be recycled for at least four times without significant loss of binding capacity to proteins. These materials show great potential to separate His-tagged protein with low-cost purification at industrial scale. |
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