Fe_3O_4@PAM@NTA-Ni^(2+) Magnetic Composite Nanoparticles for Highly Specific Separation of His-tagged Proteins  

作　　者：郭惠玲 SUN Honghao SU Zhengding HU Shuchao WANG Xiaotao 

机构地区：[1]School of Food and Pharmaceutical Engineering, Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Hubei University of Technology [2]School of Biological Engineering, Hubei Provincial Cooperative Innovation Center of Industrial Fermentation, Hubei University of Technology [3]Shool of Materials Science and Engineering, Hubei University of Technology

出　　处：《Journal of Wuhan University of Technology(Materials Science)》2018年第3期559-565,共7页武汉理工大学学报（材料科学英文版）

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);Chutian Scholars Fund Project from the Education Department of Hubei Province;Hundred Talents Program from the Organization Department of Hubei Province

摘　　要：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.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.

关 键 词：magnetic nanoparticles polyacrylamide protein separation his-tagged protein 

分 类 号：TB33[一般工业技术—材料科学与工程] TB383.1