Magnetically purification/immobilization of poly histidine-tagged proteins by PEGylated magnetic graphene oxide...

Carbon-based nanomaterials have many applications in biomedicine due to their unique mechanical, chemical, and biological properties. Among them, graphene has received special attention due to its very high specific surface area, high flexibility, and chemical stability. In this study, graphene oxide was first functionalized with amine groups (GO-NH₂) and then Fe₃O₄ nanoparticles were deposited on it using the hydrothermal method. In addition, polyethylene glycol (PEG) was attached to the magnetic graphene nanoparticles to increase their stability and solubility. Finally, PEGylated magnetic graphene nanocomposites were functionalized with nickel-nitrilotriacetic acid (NTA-Ni⁺²) to bind to the poly-histidine tag in recombinant proteins. The resulting nanocomposites (MG-PEG-NTA-Ni⁺²) were then used for magnetic immobilization and purification of recombinant β-NGF as a protein with his-tag sequence. Binding and purification were confirmed by FTIR and SDS-PAGE techniques, respectively. Importantly, differentiation of the PC12 cell line into neurons demonstrated that the purified β-NGF was fully functional. Our results suggest that MG-PEG-NTA-Ni⁺² nanocomposites may be a suitable alternative to commercial resins for rapid and specific protein immobilization and purification.