Aggregation-resistant nanozyme containing accessible magnetite nanoparticles immobilized in monodisperse-porous silica microspheres for colorimetric assay of human genomic DNA

J Colloid Interface Sci. 2019 Aug 15;550:90-98. doi: 10.1016/j.jcis.2019.04.089. Epub 2019 Apr 29.

Abstract

Bridging-induced aggregation of individual nanozyme particles by long-chain biomacromolecules causes loss of peroxidase mimetic activity for various nanozymes. When a common nanozyme (Fe3O4 nanoparticles) was used for colorimetric assay of a long-chain biomacromolecule, human genomic DNA (hgDNA, 14.000 kDa, containing 22 kilobases), peroxidase-like activity was absent owing to the irreversible aggregation of Fe3O4 nanoparticles in the presence of hgDNA. We synthesized an aggregation-resistant nanozyme containing Fe3O4 nanoparticles immobilized in 5.1 μm monodisperse-porous silica microspheres. Fe3O4 nanoparticles were immobilized in a form accessible to the substrate and large biomolecules in the solution within the monodispersed silica microspheres including both mesopores and macropores. An appreciable and stable spectrophotometric response was obtained, originating from the satisfactorily high peroxidase-like activity of the synthesized nanozyme. No aggregation was observed in the aqueous dispersion of the nanozyme in the presence of hgDNA. Large particle size, low particle number density, high surface area, and the presence of macropores were evaluated as factors contributing to the adsorption of hgDNA chains onto the synthesized nanozyme without interparticle bridge formation between the individual microspheres causing aggregation. Here, for the first time a colorimetric assay was developed based on the enhancement of peroxidase-mimetic activity of non-aggregated porous silica microspheres to determine hgDNA concentration up to 300 ng/μL. hgDNA could be also isolated with 47% yield and an equilibrium hgDNA adsorption of 19.000 ng/mg, using the same nanozyme. Hence, a material acting as a nanozyme for colorimetric determination of hgDNA was also evaluated as a magnetic, solid phase extraction sorbent for the first time.

Keywords: Biosensor; Colorimetric assay; DNA purification; Magnetic particles; Nanozyme; Peroxidase-like activity.

MeSH terms

  • Adsorption
  • Biological Assay / methods
  • Colorimetry / methods
  • DNA / analysis*
  • DNA / isolation & purification
  • Ferrosoferric Oxide / chemistry
  • Genomics / methods
  • Humans
  • Limit of Detection
  • Magnetics / methods
  • Magnetite Nanoparticles / chemistry*
  • Microspheres*
  • Particle Size
  • Peroxidase / chemistry
  • Porosity
  • Silicon Dioxide / chemistry*
  • Solid Phase Extraction / methods
  • Surface Properties

Substances

  • Magnetite Nanoparticles
  • Silicon Dioxide
  • DNA
  • Peroxidase
  • Ferrosoferric Oxide