Electrochemiluminescence Biosensor for Hyaluronidase Based on the Ru(bpy)₃²⁺ Doped SiO₂ Nanoparticles Embedded in the Hydrogel Fabricated by Hyaluronic Acid and Polyethylenimine

Abstract

Hyaluronidase (HAase), a specific enzyme of hyaluronic acid (HA), has been reported as a potential tumor biomarker in recent years. Hence, developing some simple, rapid, and sensitive methods for HAase assay is necessary. In this work, a simple and sensitive biosensor constructed by a reliable controlled release system and a mature electrochemiluminescence (ECL) analytical technique has been devised for the quantification of HAase with high efficiency and selectivity. Tris (2,2′-bipyridyl) ruthenium(II) chloride hexahydrate doped SiO2 nanoparticles (Ru@SiO2 NPs), as ECL signal probes, were trapped in the hydrogel fabricated by HA and polyethylenimine evenly and steadily. When HAase existed, the hydrogel was decomposed by HAase, and the Ru@SiO2 NPs escaped from the hydrogel into the supernate. The ECL signal produced from the supernate can be detected and used to characterize HAase concentration. The result showed a good linear relationship between ECL intensity, and HAase concentration ranged from 2 to 60 U/mL and the limit of detection was 2 U/mL. The developed controlled release ECL biosensor has been used for HAase quantification in urine samples with satisfactory results.