• 文献标题:   Magnetically purification/immobilization of poly histidine-tagged proteins by PEGylated magnetic graphene oxide nanocomposites
  • 文献类型:   Article
  • 作  者:   KHATAMINEZHAD ES, HAJIHASSAN Z, ASTARAEI FR
  • 作者关键词:   graphene oxide, polyethylene glycol, protein immobilization, magnetically protein purification
  • 出版物名称:   PROTEIN EXPRESSION PURIFICATION
  • ISSN:   1046-5928 EI 1096-0279
  • 通讯作者地址:  
  • 被引频次:   0
  • DOI:   10.1016/j.pep.2023.106264 EA MAR 2023
  • 出版年:   2023

▎ 摘  要

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 nano-composites (MG-PEG-NTA-Ni+2) were then used for magnetic immobilization and purification of recombinant 8-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 8-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.