title: Efficient purification of His-tagged protein by superparamagnetic Fe3O4/Au-ANTA-Co2+ nanoparticles.
abstract: Superparamagnetic Fe3O4/Au nanoparticles were synthesized and surface modified with mercaptopropionic acid (MPA), followed by conjugating Nα,Nα-Bis(carboxymethyl)-l-lysine hydrate (ANTA) and subsequently chelating Co(2+). The resulting Fe3O4/Au-ANTA-Co(2+) nanoparticles have an average size of 210 nm in aqueous solution, and a magnetization of 36 emu/g, endowing the magnetic nanoparticles with excellent magnetic responsivity and dispersity. The Co(2+) ions in the magnetic nanoparticle shell provide docking site for histidine, and the Fe3O4/Au-ANTA-Co(2+) nanoparticles exhibit excellent performance in binding of a His-tagged protein with a binding capacity of 74 μg/mg. The magnetic nanoparticles show highly selective purification of the His-tagged protein from Escherichia coli lysate. Therefore, the obtained Fe3O4/Au-ANTA-Co(2+) nanoparticles exhibited excellent performance in the direct separation of His-tagged protein from cell lysate.
authors.affiliation: Department of Life Science, Dezhou University, Dezhou 253023, PR China. lianyingzh@yahoo.com
authors.forename: Lianying
authors.initials: L
authors.lastname: Zhang
authors.forename: Xinjun
authors.initials: X
authors.lastname: Zhu
authors.forename: Dejie
authors.initials: D
authors.lastname: Jiao
authors.forename: Yongling
authors.initials: Y
authors.lastname: Sun
authors.forename: Hanwen
authors.initials: H
authors.lastname: Sun
mesh_terms: D002457:Cell Extracts; D003035:Cobalt / Q000737:chemistry*; D003911:Dextrans / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006046:Gold / Q000737:chemistry*; D006639:Histidine / Q000302:isolation & purification*; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry*; D060328:Magnetic Phenomena; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009842:Oligopeptides / Q000302:isolation & purification*; D010316:Particle Size; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*; D059027:Recycling; D000685:Serum Amyloid A Protein / Q000302:isolation & purification; D013057:Spectrum Analysis
chemical_list: C097388:2-N,N-bis(carboxymethyl)lysine; D002457:Cell Extracts; D003911:Dextrans; C471213:His-His-His-His-His-His; D058185:Magnetite Nanoparticles; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D000685:Serum Amyloid A Protein; D003035:Cobalt; D006639:Histidine; D006046:Gold; C065507:ferumoxides; D008239:Lysine