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 suspen...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 Ni^(2+) 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-Ni^(2+) 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.展开更多
基金Funded by the National Natural Science Foundation of China(Nos.21401051 and 51303049)Hubei Province Natural Science Foundation of China(Nos.2014CFB595 and 2014CFA080)+1 种基金Chutian Scholars Fund Project from the Education Department of Hubei ProvinceHundred Talents Program from the Organization Department of Hubei Province
文摘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 Ni^(2+) 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-Ni^(2+) 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.