Monodispersed silica nanoparticles as carrier for co-immobilization of bi-enzyme and its application for glucose...

A novel glucose sensing strategy by using bi-enzyme coated monodispered silica nanoparticles (SiO2) was proposed. The monodispered SiO2 was synthesized according to our previously reported seed-growth methods. Glucose oxidase (GOD) and horseradish peroxidase (HRP) were simultaneously covalent immobilized on the surface of SiO2 nanoparticles through the cross-linker of glutaraldehyde. The immobilized bi-enzyme remained their bioactivities well for the substrate reaction. Thus, the resultant SiO2-GOD/HRP nanocomposites could be used as catalyst for enzymatic substrate reactions in the presence of 3,3',5,5'-tetramethylbenzidine (TMB) as chromogenic reagent and glucose as substrate. The factors of affecting the catalytic activities of enzymes were optimized. Under optimal conditions, the absorbance at 450 nm in UV-visible spectra increased with the glucose concentration, which could be used for glucose detection with a linear range from 0.5 μM to 250 μM and a detection limit of 0.22 μM at a signal-to-noise ratio of 3σ. Considering the potential of making pills using this SiO2-GOD/HRP, the present strategy has good prospect in the clinic science and other fields in future.