Streptavidin-modified magnetic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) microspheres for selective isolation of bacterial DNA

摘要

The dispersion copolymerization of 2-hydroxyethyl methacrylate (HEMA) with glycidyl methacrylate (GMA) in a toluene/2-methylpropan-1-ol mixture in the presence of Fe_3O_4 nanoparticles coated with oleic acid, produced monodisperse magnetic poly(2-hydroxyethyl methacrylate-co-glycidyl methacrylate) (P(HEMA-GMA)) microspheres. Oxirane groups of the microspheres were ammonolyzed and then functionalized with streptavidin using cyanuric chloride. The final product contained 0.67 mg of streptavidin per gram of wet magnetic P(HEMA-GMA) microspheres. The microspheres were characterized by elemental analysis, scanning electron microscopy, IR, UV-VIS and atomic absorption spectroscopy. The streptavidin-modified magnetic P(HEMA-GMA) microspheres were used for immobilization of biotinylated DNA and subsequent selective isolation of target DNA from complex samples using DNA/DNA hybridisation. Based on the highly selective recognition of streptavidin with a biotin-labeled DNA probe, DNA sensor was constructed for magnetic separation of DNA from real samples.