Electrochemiluminescence Biosensor for Hyaluronidase Based on the Adjustable Electrostatic Interaction between the Surface-Charge-Controllable Nanoparticles and Negatively Charged Electrode

A novel electrochemiluminescence (ECL) biosensor for hyaluronidase (HAase) based on the adjustable electrostatic interaction between the surface-charge-controllable nanoparticles and negatively charged electrode has been devised. Hyaluronic acid (HA)-coated amino-modified ruthenium bipyridine-doped silica nanoparticles (Ru@SiO₂-NH₂@HA NPs) have been synthesized and act as ECL indicators, and the surface of this particle is negatively charged because HA contains a large amount of OH- and COO-. The strong electrostatic repulsion between the Ru@SiO₂-NH₂@HA NPs and negatively charged indium tin oxide (ITO) electrode surface leads to the detection of a low-intensity ECL signal. In the presence of HAase, the HA on the surface of the Ru@SiO₂-NH₂@HA NPs can be decomposed, and the particles can be transformed into positively charged amino-modified ruthenium bipyridine-doped silica nanoparticles (Ru@SiO₂-NH₂ NPs), which can be concentrated near the surface of the ITO electrode through electrostatic attraction, and result in the detection of an enhanced ECL signal. The ECL of the system has a good linear relationship with HAase concentration in the range of 2.0-60 U/mL, and the limit of detection was 0.37 U/mL. The designed biosensor had been applied to detect the target in real samples with satisfied results.