title: Improving the binding capacity of Ni2+ decorated porous magnetic silica spheres for histidine-rich protein separation.
abstract: Biomagnetic immobilization of histidine-rich proteins based on the single-step affinity adsorption of transition metal ions continues to be a suitable practice as a cost effective and a up scaled alternative to the to multiple-step chromatographic separations. In our previous work, we synthesised Porous Magnetic silica (PMS) spheres by one-step hydrothermal-assisted modified-stöber method. The obtained spheres were decorated with Ni(2+) and Co(2+), and evaluated for the capture of a H6-Tagged green fluorescence protein (GFP-H6) protein. The binding capacity of the obtained spheres was found to be slightly higher in the case Ni(2+) decorated PMS spheres (PMSNi). However, comparing with commercial products, the binding capacity was found to be lower than the expected. In this way, the present work is an attempt to improve the binding capacity of PMSNi to histidine-rich proteins. We find that increasing the amount of Ni(2+) onto the surface of the PMS spheres leads to an increment of the binding capacity to GFP-H6 by a factor of two. On the other hand, we explore how the size of histidine-rich protein can affect the binding capacity comparing the results of the GFP-6H to those of the His-tagged α-galactosidase (α-GLA). Finally, we demonstrate that the optimization of the magnetophoresis parameters during washing and eluting steps can lead to an additional improvement of the binding capacity.
authors.affiliation: Centro de Fisica, Universidade do Minho, 4710-057 Braga, Portugal. benelmekki@fisica.uminho.pt
authors.forename: M
authors.initials: M
authors.lastname: Benelmekki
authors.forename: C
authors.initials: C
authors.lastname: Caparros
authors.forename: E
authors.initials: E
authors.lastname: Xuriguera
authors.forename: S
authors.initials: S
authors.lastname: Lanceros-Mendez
authors.forename: E
authors.initials: E
authors.lastname: Rodriguez-Carmona
authors.forename: R
authors.initials: R
authors.lastname: Mendoza
authors.forename: J L
authors.initials: JL
authors.lastname: Corchero
authors.forename: Ll M
authors.initials: LM
authors.lastname: Martinez
mesh_terms: D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D003035:Cobalt / Q000737:chemistry; D049452:Green Fluorescent Proteins; D006639:Histidine / Q000302:isolation & purification*; D008280:Magnetics; D046529:Microscopy, Electron, Transmission; D009532:Nickel / Q000737:chemistry*; D016062:Porosity; D011506:Proteins / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry*; D013535:Suspensions; D000519:alpha-Galactosidase / Q000737:chemistry
chemical_list: D002593:Cetrimonium Compounds; D011506:Proteins; D013535:Suspensions; D049452:Green Fluorescent Proteins; D003035:Cobalt; D006639:Histidine; D012822:Silicon Dioxide; D009532:Nickel; D000519:alpha-Galactosidase; D000077286:Cetrimonium