title: Carbonaceous materials passivation on amine functionalized magnetic nanoparticles and its application for metal affinity isolation of recombinant protein.
abstract: Magnetic nanoparticles (MNPs) with an amine functionalized surface (MH) were passivated with carbonaceous materials (MH@C) by carbonization of glucose under hydrothermal reaction conditions. The carboxylate groups in carbonaceous shell could be enriched to 0.285 mmol/g when acrylic acid was added as a functional monomer in the carbonization reaction (MH@C-Ac). The carbonaceous shell not only protected the magnetic core from acidic erosion but also showed a high adsorption capacity toward Ni(2+) ion. The Ni(2+) ion complexed on MH@C and MH@C-Ac could specifically isolate 6×His tagged recombinant proteins from crude bacterial extracts via metal affinity interaction. The superparamagnetic property facilitates the easy retrieval of the carbonaceous material passivated MNPs from the viscous proteins solutions. Recombinant green fluorescence protein (GFP) and hyaluronic acid (HA) lyase of 9.4 mg and 2.3 mg could be isolated by 1 g of MH@C-Ac-Ni, respectively.
authors.affiliation: Department of Chemical Engineering, National Taiwan University of Science and Technology, 43 Keelung Rd Sec. 4, Taipei 106, Taiwan.
authors.forename: Iryanti F
authors.initials: IF
authors.lastname: Nata
authors.forename: Nermeen S
authors.initials: NS
authors.lastname: El-Safory
authors.forename: Cheng-Kang
authors.initials: CK
authors.lastname: Lee
mesh_terms: D000588:Amines / Q000737:chemistry*; D002244:Carbon / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D005290:Ferric Compounds / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D006821:Hyaluronoglucosaminidase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D008670:Metals / Q000737:chemistry*; D009532:Nickel / Q000737:chemistry; D009842:Oligopeptides / Q000737:chemistry; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000588:Amines; D002712:Chlorides; D005290:Ferric Compounds; C471213:His-His-His-His-His-His; D058185:Magnetite Nanoparticles; D008670:Metals; D009842:Oligopeptides; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D006639:Histidine; D002244:Carbon; D009532:Nickel; D006821:Hyaluronoglucosaminidase; C024555:ferric chloride