摘要 :
OBJECTIVES:Peroxidase-oxidase mimetic activity materials are important for developing new commercial assays. MATERIALS and METHODS:A new staged-shape templated hydrolysis and condensation protocol was developed for the synthesis o...
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OBJECTIVES:Peroxidase-oxidase mimetic activity materials are important for developing new commercial assays. MATERIALS and METHODS:A new staged-shape templated hydrolysis and condensation protocol was developed for the synthesis of monodisperse-porous metal oxide microspheres.The magnetic forms were synthesized with the accessible magnetic hematite (Fe3O4 ) nanoparticles tightly immobilized within the porous interiors of microspheres.Magnetic SiO2 microspheres and plain CeO2 microspheres exhibited peroxidase mimetic activity while oxidase-mimetic activity was obtained with the plain MnO2 microspheres. RESULTS:3,3’,5,5’-tetramethylbenzidine (TMB) and o-phenylenediamine (OPDA) were used as the synthetic substrates providing products via oxidase/ peroxidase-like activities of corresponding microspheres, whose formation kinetics were followed by UV-Vis spectroscopy or fluorescence spectroscopy. The interaction of plain and ligand-attached forms of porous metal oxide microspheres with the biomolecules resulted in a change in the peroxidaseoxidase mimetic activity which was proportional to the biomolecule concentration in the interaction medium, due to the selective adsorption of biomolecule onto the surface of microspheres. CONCLUSIONS:The concentration of ascorbic acid was determined via oxidaselike activity of plain, porous MnO2 microspheres, using both colorimetric and fluorometric protocols. The ligand attached forms of plain SiO2 microspheres were evaluated for developing a colorimetric assay for the determination of histidine tagged protein concentration.We believe that the methodology exemplified here will be effectively used for developing new commercial assays for the estimation of concentrations of various agents in biological samples. The research was supported by Hacettepe University Scientific Research Projects Coordination Unit contract numbered as FBA-2019-17337, entitled ‘’ Development of a peroxidase-microzyme-based biosensor for protein determination’’. The financial support provided is gratefully acknowledged.
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摘要 :
OBJECTIVES: This study was to test the peroxidase activity of the monodispers_x0002_porous silica microspheres.The use of synthesized microspheres as a support material reveals a linear increase in peroxidase activity with increas...
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OBJECTIVES: This study was to test the peroxidase activity of the monodispers_x0002_porous silica microspheres.The use of synthesized microspheres as a support material reveals a linear increase in peroxidase activity with increased concentration of genomic DNA.The results facilitate the development of a new diagnostic kit.MATERIALS and METHODS: The monodisperse-porous silica microspheres were 5 mm in size and with a coefficient of variation for size distribution less than 5%.The morphological investigation performed by SEM showed that the surface of microspheres was porous.The devleoped synthetic protocol of the magnetic silica microspheres allowed strong immobilization of magnetic Fe3O4 nanoparticles on the surface of the pore found within the microspheres.The presence of tightly immobilized Fe3O4 nanoparticles having an ability to interact with the enviroment should be probably the main reason of peroxidase-like activity of magnetic silica microspheres.RESULTS: In order to test the peroxidase-like activity of magnetic silica microspheres, tetramethylbenzidine (TMB) was used as the substrate.The effect of concentration of magnetic silica microspheres on the peroxidaselike activity was investigated in TE buffer medium using TMB as the synthetic substrate and a linear increase in the peroxidase-like activity with the increasing microsphere concentration.Besides, the effect of human genomic DNA concentration on the peroxidase-like activity of magnetic silica microspheres was investigated.The results showed that a linear increase occurred in the activity with the increasing DNA concentration.CONCLUSIONS: The results of presented study also allow the development of new diagnostic methods worked based on peroxidase-activity.The magnetic silica microspheres have a significant potential as support material for various biological molecules.Hence, the use of magnetic silica microspheres with this property instead of an expensive enzyme-peroxidase may provide serious advantages in the studies on the development of diagnostic test kits.
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