Fabrication Of Functionalized Magnetic Nanoparticles For Protein Separation Methodology Research

Protein is an important component of the living organisms. In recent years, the research of protein separation technology has been widely concerned. The development of stable, simple, efficient protein adsorbents materials has become an important research topic in the field of biochemistry. Nanomaterials play an important role in the field of biological separation, as a novel functional nanomaterials, magnetic nanoparticles have received more and more attentions and show great potential in the application of biological macromolecules separation because of their simple preparation process, unique magnetic response property, high surface-to-volume ratios and good biocompatibility, etc. Molecular imprinting is a technique to prepare the molecularly imprinted polymers (MIPs) with recognition sites having a predetermined selectivity for given molecules. At present, the molecularly imprinting technique is a mature method for the separation and analysis of small molecule compounds. As the intensive study of proteomics, molecular imprinting technique has also made some progress in the application of protein separation, enrichment and purification.Based on the previous research, in this paper, proteins as study targets, we prepared protein molecular imprinted polymers modified magnetic composite materials with specific selectivity and high load amount in combination with magnetic nanomaterials and molecular imprinting technique. The adsorption performances of the novel magnetic nanoparticles as an adsorbents to target protein were investigated. The main content was as follows:First, the magnetic nanoparticles Fe3O4 with uniform size were synthesized using a modified solvothermal reaction. The surface of the Fe3O4 nanospheres was coated by a thin silicashell to form core-shell Fe3O4@SiO2 nanospheres with good dispersibility and stablility using a sol-gel process. Afterwards, an alkylation reaction was carried out to introduce vinyl groups modified nanoparticles (vinyl-Fe3O4@SiO2). Further hemoglobin was chosen as the template and the vinyl-Fe3O4@SiO2 nanospheres as the supports, surface copolymerization of acryl amide (AAm, functionalized monomer) and N,N’-methylenebisacrylamide (MBA, cross-linker) was performed in aqueous media. In addition, MIPs thin films coated magnetic nanospheres (Fe3O4@SiO2@Hb-MIPs) with a core-shell structure were eventually obtained after the removal of the embedded templates and non-imprinted magnetic nanospheres (Fe3O4@SiO2@NIPs) without the template protein. Finally, the separation of hemoglobin was systematically investigated using the hemoglobin molecule imprinted composite magnetic beads as adsorbent media materials, and compared with the non-imprinted materials.